Enantiomerically purified gper agonist for use in treating disease states and conditions

ABSTRACT

The present disclosure provides 1) an enantiomerically purified compound SRR G-1, or a derivative thereof, including specific crystal forms, salts and co-crystals that modulates G protein-coupled estrogen receptor activity, 2) pharmaceutical and cosmetic compositions comprising an enantiomerically purified SRR G-1, or a derivative thereof, and 3) methods of treating or preventing disease states and conditions and cosmetic conditions mediated through these receptors and related methods thereof in humans and animals.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 16/518,516 filed Jul. 22, 2019 which claims priority from U.S.Provisional Application No. 62/701,726 filed Jul. 21, 2018, each ofwhich is hereby incorporated by reference in its entirety.

GOVERNMENT INTERESTS

This invention was made with United States Government support underGrant No. 2R44CA228695-02 awarded by the National Cancer Institute ofthe National Institutes of Health. The United States Government hascertain rights in the invention.

SUMMARY

Embodiments of the present invention relate to an enantiomericallypurified agonist of the G-protein coupled estrogen receptor (GPER),pharmaceutical compositions comprising an enantiomerically purified SRRG-1, or a derivative thereof, and methods of treating disease states andconditions in subjects in need thereof, and methods of treating diseasestates and conditions mediated through GPER receptors.

Estrogens mediate multiple complex physiological responses throughoutthe body. The responses are in turn mediated through the binding ofestrogen to receptors. The classical receptors bind steroids, such asestrogen, and are soluble cytoplasmic/nuclear proteins that function astranscription factors. These receptors are known as estrogen receptoralpha and beta (two closely related proteins) that mediatetranscriptional activity. GPER is a 7-transmembrane G protein-coupledreceptor that also binds to estrogen with high affinity (K_(d) ^(˜)6 nM)and mediates rapid cellular responses including cyclic adenosinemonophosphate signaling, calcium mobilization and phosphatidylinositol3,4,5 trisphosphate production.

Diseases whose development, progression, and or response to therapy, maybe influenced by endogenous, and/or pharmacologic activation of GPERsignaling include cancer (including the prevention of cancer, preventionof the reoccurrence of cancer, and the inhibition of the progression ofcancer; and particularly melanoma, pancreatic, lymphomas, uvealmelanoma, non-small cell lung cancer, breast, reproductive and otherhormone-dependent cancers, leukemia, colon cancer, prostate, bladdercancer), reproductive (genito-urological) including endometritis,prostatitis, polycystic ovarian syndrome, bladder control,hormone-related disorders, hearing disorders, cardiovascular conditionsincluding hot flashes and profuse sweating, hypertension, stroke,obesity, diabetes, osteoporosis, hematologic diseases, vascular diseasesor conditions such as venous thrombosis, atherosclerosis, among numerousothers and disorders of the central and peripheral nervous system,including depression, insomnia, anxiety, neuropathy, multiple sclerosis,neurodegenerative disorders such as Parkinson's disease and Alzheimer'sdisease, as well as inflammatory bowel disease, Crohn's disease, coeliac(celiac) disease and related disorders of the intestine.

DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an atomic displacement ellipsoid drawing of SSR G-1dichloromethane solvate.

FIG. 2 shows a packing diagram viewed along the crystallographic a axis.

FIG. 3 shows a packing diagram viewed along the crystallographic b axis.

FIG. 4 shows a packing diagram viewed along the crystallographic c axis.

FIG. 5 shows the one-dimensional hydrogen bond network.

FIG. 6 show SRR G-1 with labeled chiral centers.

FIG. 7 shows a calculated XRPD pattern of SRR G-1 dichloromethanesolvate, generated from the single crystal structure.

FIG. 8 shows the Atomic displacement ellipsoid diagram of SRR G-1 FormA.

FIG. 9 shows the Calculated and experimental XRPD patterns for SRR G-1Form A.

FIG. 10 shows the XRPD patterns for SRR G-1 Forms A, B, and C.

FIG. 11A and FIG. 11B show the Thermograms for SRR G-1 Form A.

FIG. 12 shows the DVS isotherm for SRR G-1 Form A.

FIG. 13 shows the Atomic displacement ellipsoid diagram of SRR G-1 FormB.

FIG. 14 shows the Calculated and experimental XRPD patterns for SRR G-1Form B.

FIG. 15A and FIG. 15B show the Thermograms for SRR G-1 Form B.

FIG. 16 shows the DSC thermogram for mixture SRR G-1 Forms B and C.

FIG. 17 shows the XRPD indexing results for SRR G-1 Form C.

FIG. 18 shows the DSC thermogram for SRR G-1 Form C.

FIG. 19 shows the XRPD overlay of residual solids after pH solubilitytest (II/II).

FIG. 20 shows the XRPD overlay of residual solids after pH solubilitytest (II/II).

FIG. 21 shows the Solubility of SRR G-1 freebase in bio-relevant media.

FIG. 22 shows the XRPD overlay of SRR G-1 after solubility test in SGF.

FIG. 23 shows the XRPD overlay of SRR G-1 after solubility test inFaSSIF.

FIG. 24 shows the XRPD overlay of SRR G-1 after solubility test inFeSSIF.

FIG. 25 shows the XRPD patterns of SRR G-1 salts.

FIG. 26 shows the Atomic displacement ellipsoid diagram of SRR G-1Besylate Form A.

FIG. 27 shows the Calculated and experimental XRPD patterns for SRR G-1Besylate Form A.

FIG. 28A and FIG. 28B show the Thermograms for SRR G-1 Besylate Form A.

FIG. 29 shows the Indexing results for SRR G-1 Camsylate Form A.

FIG. 30 shows the XRPD pattern for SRR G-1 Camsylate Form A shown from 5to 19° (2θ).

FIG. 31A and FIG. 31B show the Thermograms for SRR G-1 Camsylate Form A.

FIG. 32 shows the Indexing results for SRR G-1 Napsylate Form A.

FIG. 33A and FIG. 33B shows the Thermograms for SRR G-1 Napsylate FormA.

FIG. 34 illustrates the results of a proliferation assay using YUMM1.7melanoma cells. In this assay, the cells were treated with 500 nM of theracemic mixture (G-1), or the single enantiomers of G-1 SRR G-1, and RSSG-1. The dotted line indicates starting cell population number. n=5replicates per group. *denotes p<0.05, error bars=±s.d.

FIG. 35 shows the Plasma Concentration of SRR G-1 in the Rat Dosed withSRR G-1 Free Base.

FIG. 36 shows the Plasma Concentration of SRR G-1 in the Rat Dosed withSRR G-1 Besylate.

FIG. 37 shows the Plasma Concentration of SRR G-1 in the Rat Dosed withSRR G-1 Napsylate.

FIG. 38 shows the Comparison of Plasma Concentrations of SRR G-1 in theRat Dosed with SRR G-1 Free Base, SRR G-1 Besylate, and SRR G-1Napsylate.

DETAILED DESCRIPTION Definitions

As used herein, the terms below have the meanings indicated.

Before the present compounds, compositions and methods are described, itis to be understood that this invention is not limited to the particularprocesses, formulations, compound, compositions, or methodologiesdescribed, as these may vary. It is also to be understood that theterminology used in the description is for the purpose of describing theparticular versions or embodiments only, and is not intended to limitthe scope of embodiments herein which will be limited only by theappended claims. Unless defined otherwise, all technical and scientificterms used herein have the same meanings as commonly understood by oneof ordinary skill in the art. Although any methods and materials similaror equivalent to those described herein can be used in the practice ortesting of embodiments herein, the preferred methods, devices, andmaterials are now described. All publications mentioned herein areincorporated by reference in their entirety. Nothing herein is to beconstrued as an admission that embodiments herein are not entitled toantedate such disclosure by virtue of prior invention.

It must also be noted that as used herein and in the appended claims,the singular forms “a,” “an,” and “the” include plural reference unlessthe context clearly dictates otherwise.

As used herein, the term “about” means plus or minus 20% of thenumerical value of the number with which it is being used. Therefore,about 50% means in the range of 40%-60%.

In embodiments or claims where the term “comprising” is used as thetransition phrase, such embodiments can also be envisioned withreplacement of the term “comprising” with the terms “consisting of” or“consisting essentially of.”

As used herein, the term “consists of” or “consisting of” means that thecompound, composition, formulation or the method includes only theelements, steps, or ingredients specifically recited in the particularclaimed embodiment or claim.

As used herein, the term “consisting essentially of” or “consistsessentially of” means that the compound, composition, formulation or themethod includes only the elements, steps or ingredients specificallyrecited in the particular claimed embodiment or claim and may optionallyinclude additional elements, steps or ingredients that do not materiallyaffect the basic and novel characteristics of the particular embodimentor claim. For example, the only active ingredient(s) in the formulationor method that treats the specified condition (e.g., cancer and/orobesity) is the specifically recited therapeutic(s) in the particularembodiment or claim.

As used herein, the term “a derivative thereof” refers to any molecularform of the compound it references, including, but not limited to, asalt thereof, a pharmaceutically acceptable salt thereof, an esterthereof, a free base thereof, a solvate thereof, a hydrate thereof, anN-oxide thereof, a clathrate thereof, a prodrug thereof, an isotopethereof (e.g., tritium, deuterium), a co-crystal thereof, and anycombination of the foregoing.

Asymmetric centers exist in the compounds disclosed herein. Thesecenters are designated by the symbols “R” or “S,” depending on theconfiguration of substituents around the chiral carbon atom. It shouldbe understood that the invention encompasses all stereochemical isomericforms, including diastereomeric, enantiomeric, and epimeric forms, andmixtures thereof. Individual stereoisomers of compounds can be preparedsynthetically from commercially available starting materials whichcontain chiral centers or by preparation of mixtures of enantiomericproducts followed by separation such as conversion to a mixture ofdiastereomers followed by separation or recrystallization,chromatographic techniques, direct separation of enantiomers on chiralchromatographic columns, or any other appropriate method known in theart. Starting compounds of particular stereochemistry are eithercommercially available or can be made and resolved by techniques knownin the art. Additionally, the compounds disclosed herein may exist asgeometric isomers. The present invention includes all cis, trans, syn,anti, entgegen (E), and zusammen (Z) isomers as well as the appropriatemixtures thereof. Additionally, compounds may exist as tautomers; alltautomeric isomers are provided by this invention. Additionally, thecompounds disclosed herein may exist in unsolvated as well as solvatedforms with pharmaceutically acceptable solvents such as water, ethanol,and the like. In general, the solvated forms are considered equivalentto the unsolvated forms.

As used herein, the term “chiral purity” and “enantiomeric excess” (ee)are interchangeable and may refer to the measurement of the absolutedifference between the mole fraction of each enantiomer and is mostoften expressed as a percentage. % Enantiomeric excess may be determinedby the formula:

% ee=|A−B|×100

Where A and B are the respective mole fractions of the enantiomers in amixture such that A+B=1. A racemic mixture has an enantiomeric excess of0%, while a single completely pure enantiomer has an enantiomeric excessof 100%. As an example, a sample with 70% of R isomer and 30% of S willhave an enantiomeric excess of 40%. This can also be thought of as amixture of 40% pure R with 60% of a racemic mixture (which contributes30% R and 30% S to the overall composition).

The term “substantially free” as used herein, alone or in combination,refers to the absence of isomers within the limits of quantitation ofanalytical methods such as nuclear magnetic resonance (NMR), gaschromatography/mass spectroscopy (GC/MS), high performance liquidchromatography (HPLC), circular dichroism (CD), or other methods ofchemical analysis.

“Pharmaceutically acceptable salt” is meant to indicate those salts orco-crystals which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of a patient without unduetoxicity, irritation, allergic response and the like, and arecommensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, Berge et al.(1977) J. Pharm. Sciences, Vol. 6, 1-19, describes representativepharmaceutically acceptable salts in detail. A pharmaceutical acceptable“salt” is any acid addition salt or co-crystal, preferably apharmaceutically acceptable acid addition salt or co-crystal, including,but not limited to, halogenic acid salts such as hydrobromic,hydrochloric, hydrofloric and hydroiodic acid salt; an inorganic acidsalt such as, for example, nitric, perchloric, sulfuric and phosphoricacid salt; an organic acid salt such as, for example, sulfonic acidsalts (methanesulfonic, trifluoromethan sulfonic, ethanesulfonic,benzenesulfonic or p-toluenesufonic), acetic, malic, fumaric, succinic,citric, benzonic gluconic, lactic, mandelic, mucic, pamoic, pantothenic,oxalic and maleic acid salts; and an amino acid salt such as aspartic orglutamic acid salt, benzenesulfonic, (+)-(1S)-camphor-10-sulfonic,ethane-1,2-disulfonic, hydrochloric, methanesulfonic,naphthalene-2-sulfonic, naphthalene-1,5-disulfonic, sulfuric, andp-toluenesulfonic acid. The pharmaceutically acceptable salt may be amono- or di-acid addition salt, such as a di-hydrohalogic, di-sulfuric,di-phosphoric or di-organic acid salt. The pharmaceutically acceptablesalt is used as a chiral or achiral reagent which is not required to beselected on the basis of any expected or known preference for theinteraction with or precipitation of a specific optical isomer of theproducts of this disclosure.

The term “therapeutically acceptable salt,” as used herein, representssalts or co-crystals or zwitterionic forms of the compounds disclosedherein which are water or oil-soluble or dispersible and therapeuticallyacceptable as defined herein. The salts can be prepared during the finalisolation and purification of the compounds or separately by reactingthe appropriate compound in the form of the free base with a suitableacid or by substituting one salt for an therapeutic acceptable salt.Representative acid addition salts include acetate, adipate, alginate,L-ascorbate, aspartate, benzoate, benzenesulfonate (besylate),bisulfate, butyrate, camphorate, camphorsulfonate, citrate, digluconate,formate, fumarate, gentisate, glutarate, glycerophosphate, glycolate,hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isethionate),lactate, maleate, malonate, DL-mandelate, mesitylenesulfonate,methanesulfonate, naphthylenesulfonate, nicotinate,2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate,3-phenylproprionate, phosphonate, picrate, pivalate, propionate,pyroglutamate, succinate, sulfonate, tartrate, L-tartrate,trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate,para-toluenesulfonate (p-tosylate), and undecanoate. Also, basic groupsin the compounds disclosed herein can be quaternized with methyl, ethyl,propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl,dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and sterylchlorides, bromides, and iodides; and benzyl and phenethyl bromides.Examples of acids which can be employed to form therapeuticallyacceptable addition salts include inorganic acids such as hydrochloric,hydrobromic, sulfuric, and phosphoric, and organic acids such as oxalic,maleic, succinic, and citric. Hence, the present invention contemplatessodium, potassium, magnesium, and calcium salts of the compoundsdisclosed herein, and the like.

As used herein, the term “patient” and “subject” are interchangeable andmay be taken to mean any living organism, which may be treated withcompounds of the present invention. As such, the terms “patient” and“subject” may include, but is not limited to, any non-human mammal,primate or human. In some embodiments, the “patient” or “subject” is anadult, child, infant, or fetus. In some embodiments, the “patient” or“subject” is a human. In some embodiments, the “patient” or “subject” isa mammal, such as mice, rats, other rodents, rabbits, dogs, cats, swine,cattle, sheep, horses, primates, or humans.

The terms “therapeutically effective amount” or “therapeutic dose” isused herein are interchangeable and may refer to the amount of an activeagent or pharmaceutical compound or composition that elicits a clinical,biological or medicinal response in a tissue, system, animal, individualor human that is being sought by a researcher, veterinarian, medicaldoctor or other clinical professional. A clinical, biological or medicalresponse may include, for example, one or more of the following: (1)preventing a disease, condition or disorder in an individual that may bepredisposed to the disease, condition or disorder but does not yetexperience or display pathology or symptoms of the disease, condition ordisorder, (2) inhibiting a disease, condition or disorder in anindividual that is experiencing or displaying the pathology or symptomsof the disease, condition or disorder or arresting further developmentof the pathology and/or symptoms of the disease, condition or disorder,and (3) ameliorating a disease, condition or disorder in an individualthat is experiencing or exhibiting the pathology or symptoms of thedisease, condition or disorder or reversing the pathology and/orsymptoms experienced or exhibited by the individual.

The terms “administer,” “administering” or “administration” as usedherein refer to either directly administering a compound orpharmaceutically acceptable salt of the compound or a composition to asubject.

The term “treating” may be taken to mean prophylaxis of a specificdisorder, disease or condition, alleviation of the symptoms associatedwith a specific disorder, disease or condition and/or prevention of thesymptoms associated with a specific disorder, disease or condition. Insome embodiments, the term refers to slowing the progression of thedisorder, disease or condition or alleviating the symptoms associatedwith the specific disorder, disease or condition. In some embodiments,the term refers to alleviating the symptoms associated with the specificdisorder, disease or condition. In some embodiments, the term refers toalleviating the symptoms associated with the specific disorder, diseaseor condition. In some embodiments, the term refers to restoring functionwhich was impaired or lost due to a specific disorder, disorder orcondition.

The term “preventing” may be taken to mean to prevent a specificdisorder, disease or condition and/or prevent the reoccurrence of aspecific disorder, disease or condition.

The term “unit dosage form” refers to physically discrete units suitableas a unitary dosage for human subjects and other animals, each unitcontaining a predetermined quantity of active material calculated toproduce the desired therapeutic effect, in association with a suitablepharmaceutical excipient.

The term “disease” as used herein is intended to be generallysynonymous, and is used interchangeably with, the terms “disorder,”“syndrome,” and “condition” (as in medical condition), in that allreflect an abnormal condition of the human or animal body or of one ofits parts that impairs normal functioning, is typically manifested bydistinguishing signs and symptoms, and causes the human or animal tohave a reduced duration or quality of life.

The term “combination therapy” means the administration of two or moretherapeutic agents to treat a medical condition or disorder described inthe present disclosure. Such administration encompassesco-administration of these therapeutic agents in a substantiallysimultaneous manner, such as in a single capsule, or dosagepresentation, having a fixed ratio of active ingredients or in multiple,separate capsules for each active ingredient. In addition, suchadministration also encompasses use of each type of therapeutic agent ina sequential manner in the same patient, with delivery of the individualtherapeutics separated by 1-24 hours, 1-7 days, or 1 or more weeks. Ineither case, the treatment regimen will provide beneficial effects ofthe drug combination in treating the conditions or disorders describedherein.

Compounds

Many organic compounds exist in optically active forms, i.e. they havethe ability to rotate the plane of plane polarized light. In describingan optically active compound, the prefixes R and S are used to denotethe absolute configuration of the molecule about its chiral center(s).For a given chemical structure, these compounds, called stereoisomers,are identical except that they are mirror images of one another. Aspecific stereoisomer may also be referred to as an enantiomer, and amixture of such isomers is often called an enantiomeric or racemicmixture.

Stereochemical purity is of importance in the field of pharmaceuticals,where 8 of the 10 most prescribed drugs exhibit chirality. A case inpoint is provided by the S-enantiomer of the β-adrenergic blockingagent, propranolol, which is known to be 100 times more potent than theR-enantiomer.

Embodiments of the present invention encompass compounds comprisingenantiomerically purified G-1 and methods of use in the treatment ofdiseases. G-1 is a racemic mixture of the enantiomers1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one(henceforth referred to as “SRR G-1” or “LNS8801”) and 1-((3aR,4S,9bS)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one(henceforth referred to as “RSS G-1” or “LNS8812”).

Enantiomerically purified G-1 has been purified in favor of its1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-oneenantiomer over the corresponding1-((3aR,4S,9bS)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-oneenantiomer. Unless specifically described, SRR G1, or a derivativethereof includes, any physical form, including an amorphous form or anycrystalline solid forms such as A, B, C or combinations thereof.

In certain embodiments, the compound of1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one(also referred to as “SRR G-1”), or a derivative thereof, has a chiralpurity of about 90% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 91% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 92% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 93% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 94% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 95% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 96% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 97% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 97.5% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 98% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.1% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99.2% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.3% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99.4% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.5% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99.6% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.7% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99.75% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.8% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99.9% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.91% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99.92% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.93% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99.94% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.95% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99.96% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.97% or greater. Incertain embodiments SRR G-1, or a derivative thereof, has a chiralpurity of about 99.98% or greater. In certain embodiments SRR G-1, or aderivative thereof, has a chiral purity of about 99.99% or greater. Incertain embodiments SRR G-1, or a derivative thereof, is free of itsopposite enantiomer within the limits of quantification. In certainembodiments SRR G-1, or a derivative thereof, is substantially free ofits opposite enantiomer.

In any of the embodiments of SRR G-1 described herein, wherein thecompound is crystalline as evidenced by XRPD analysis or amorphous asevidenced by XRPD analysis or a mixture of crystalline and amorphousmaterial.

In any of the embodiments of SRR G-1 described herein, the form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one,is selected from crystalline Form A that is characterized by an XRPDpattern of FIG. 10, crystalline Form B that is characterized by an XRPDpattern of FIG. 10, crystalline Form C that is characterized by an XRPDpattern of FIG. 10, amorphous, or combinations thereof.

In any of the embodiments of SRR G-1 described herein, the crystallineform of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one,is selected from crystalline Form A that is characterized by an XRPDpattern of FIG. 10, crystalline Form B that is characterized by an XRPDpattern of FIG. 10, crystalline Form C that is characterized by an XRPDpattern of FIG. 10, or combinations thereof.

In certain embodiments, the crystalline form of1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one,is crystalline Form A that is characterized by an XRPD pattern of FIG.10.

In any of the embodiments of SRR G-1 described herein, wherein the formof 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one,is selected from crystalline Form A that is characterized by an XRPDpattern having peaks expressed in degrees 2θ (±0.20) at about 5.75,about 20.54, about 20.71, about 21.25, and about 21.86; crystalline FormB that is characterized by an XRPD pattern having peaks expressed indegrees 2θ (±0.20) at about 13.98, about 15.44, about 19.67, about21.55, and about 22.05; crystalline Form C that is characterized by anXRPD pattern having peaks expressed in degrees 2θ (±0.20) at about10.73, about 12.77, about 13.49, about 16.09, and about 20.60;amorphous; or combinations thereof.

In any of the embodiments of SRR G-1 described herein, wherein thecrystalline form of1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one,is selected from crystalline Form A that is characterized by an XRPDpattern having peaks expressed in degrees 2θ (±0.20) at about 5.75,about 20.54, about 20.71, about 21.25, and about 21.86; crystalline FormB that is characterized by an XRPD pattern having peaks expressed indegrees 2θ (±0.20) at about 13.98, about 15.44, about 19.67, about21.55, and about 22.05; crystalline Form C that is characterized by anXRPD pattern having peaks expressed in degrees 2θ (±0.20) at about10.73, about 12.77, about 13.49, about 16.09, and about 20.60; orcombinations thereof.

In certain embodiments, the crystalline form of1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one,is crystalline Form A that is characterized by an XRPD pattern havingpeaks expressed in degrees 2θ (±0.20) at about 5.75, about 20.54, about20.71, about 21.25, and about 21.86. In certain embodiments, crystallineForm A that is further characterized by an XRPD pattern having peaksexpressed in degrees 2θ (±0.20) at about 5.75, about, 9.56, about 10.53,about 17.03, about 20.54, about 20.71, about 21.25, about 21.86, about24.67, and about 28.06. In certain embodiments, crystalline Form A thatis further characterized by an XRPD pattern having peaks expressed indegrees 2θ (±0.20) at about 5.75, about, 9.56, about 10.53, about 10.81,about 13.02, about 14.66, about 14.79, about 16.23, about 17.03, about20.54, about 20.71, about 21.25, about 21.86, about 24.67, and about28.06.

In certain embodiments, the crystalline form of1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one,is crystalline Form B that is characterized by an XRPD pattern havingpeaks expressed in degrees 2θ (±0.20) at about 13.98, about 15.44, about19.67, about 21.55, and about 22.05. In certain embodiments, crystallineForm B that is further characterized by an XRPD pattern having peaksexpressed in degrees 2θ (±0.20) at about 13.98, about 14.19, about15.44, about 19.67, about 20.82, about 21.55, about 22.05, about 24.65,about 26.18, and about 28.18. In certain embodiments, crystalline Form Bthat is further characterized by an XRPD pattern having peaks expressedin degrees 2θ (±0.20) at about 7.60, about 9.71, about 13.98, about14.19, about 15.44, about 18.61, about 19.67, about 20.82, about 21.55,about 22.05, about 24.65, about 26.18, and about 28.18.

In certain embodiments, the crystalline form of1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one,is crystalline Form C that is characterized by an XRPD pattern havingpeaks expressed in degrees 2θ (±0.20) at about 10.73, about 12.77, about13.49, about 16.09, and about 20.60. In certain embodiments, crystallineForm C the is further characterized by an XRPD pattern having peaksexpressed in degrees 2θ (±0.20) at about 7.69, about 8.62, about 10.73,about 12.77, about 13.49, about 16.09, about 19.86, about 20.60, about22.05, and about 22.98.

In any of the embodiments of SRR G-1 described herein, or a derivativethereof, the derivative thereof is a salt or co-crystal.

In any of the embodiments of SRR G-1 described herein, or a derivativethereof, the derivative thereof is selected from salts or co-crystalsformed with benzenesulfonic acid, with (+)-(1S)-camphor-10-sulfonicacid, with ethane-1,2-disulfonic acid, with hydrochloric acid, withmethanesulfonic acid, with naphthalene-2-sulfonic acid, withnaphthalene-1,5-disulfonic acid, with sulfuric acid, withp-toluenesulfonic acid, or combinations thereof.

In certain embodiments, the derivative thereof is a salt or co-crystalformed with benzenesulfonic acid.

In certain embodiments, the derivative thereof is a salt or co-crystalformed with (+)-(1S)-camphor-10-sulfonic acid.

In certain embodiments, the derivative thereof is a salt or co-crystalformed with naphthalene-2-sulfonic acid.

In certain embodiments, the derivative thereof is a salt or co-crystalformed with benzenesulfonic acid and is characterized by an XRPD patternhaving peaks expressed in degrees 2θ (±0.20) at about 4.26, about 6.51,about 6.71, about 16.86, about 18.92, about 19.99, about 20.29, about20.75, about 21.46, about 22.06, about 22.12, and about 23.99.

In certain embodiments, the derivative thereof is a salt or co-crystalformed with (+)-(1S)-camphor-10-sulfonic acid and is characterized by anXRPD pattern having peaks expressed in degrees 2θ (±0.20) at about 5.97,about 11.98, about 12.69, about 13.41, about 16.23, about 17.79, about18.03, about 18.77, and about 19.69.

In certain embodiments, the derivative thereof is a salt or co-crystalformed with naphthalene-2-sulfonic acid and is characterized by an XRPDpattern having peaks expressed in degrees 2θ (±0.20) at about 6.17,about 12.63, about 12.84, about 13.75, about 14.39, about 16.79, about17.07, about 17.64, about 19.22, about 19.44, about 20.43, about 21.26,about 21.78, about 22.60, about 23.38, about 26.07, and about 27.63.

In any of the embodiments of SRR G-1 described herein, SRR G-1, or aderivative thereof, and at a concentration of 500 nM has about a 2.5fold or greater increase in inhibition of cell growth in a YUMM1.7 4 daygrowth assay as compared to a racemic mixture of SRR G-1 and itsopposite enantiomer. In certain embodiments, SRR G-1, or a derivativethereof, has about a 3 fold or greater increase in inhibition of cellgrowth. In certain embodiments, SRR G-1, or a derivative thereof, hasabout a 3.5 fold or greater increase in inhibition of cell growth. Incertain embodiments, SRR G-1, or a derivative thereof, has about a 4fold or greater increase in inhibition of cell growth. In certainembodiments, SRR G-1, or a derivative thereof, has about a 4.5 fold orgreater increase in inhibition of cell growth. In certain embodiments,SRR G-1, or a derivative thereof, has about a 5 fold or greater increasein inhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 5.5 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 6 fold or greater increase in inhibitionof cell growth. In certain embodiments, SRR G-1, or a derivativethereof, has about a 6.5 fold or greater increase in inhibition of cellgrowth. In certain embodiments, SRR G-1, or a derivative thereof, hasabout a 7 fold or greater increase in inhibition of cell growth. Incertain embodiments, SRR G-1, or a derivative thereof, has about a 7.5fold or greater increase in inhibition of cell growth. In certainembodiments, SRR G-1, or a derivative thereof, has about a 8 fold orgreater increase in inhibition of cell growth. In certain embodiments,SRR G-1, or a derivative thereof, has about a 8.5 fold or greaterincrease in inhibition of cell growth. In certain embodiments, SRR G-1,or a derivative thereof, has about a 9 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 9.5 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 10 fold or greater increase ininhibition of cell growth. Or a range between any two of these values.

In certain embodiments the compound of SRR G-1, or a derivative thereof,substantially free of its opposite enantiomer and at a concentration of500 nM has about a 7.8 fold or greater increase in inhibition of cellgrowth in a YUMM1.7 4 day growth assay as compared to a racemic mixtureof SRR G-1 and its opposite enantiomer.

In any of the embodiments of SRR G-1 described herein, SRR G-1, or aderivative thereof, and at a concentration of 500 nM has about a 5 foldor greater increase in inhibition of cell growth in a YUMM1.7 4 daygrowth assay as compared the opposite enantiomer of SRR G-1, or aderivative thereof. In certain embodiments, SRR G-1, or a derivativethereof, has about a 10 fold or greater increase in inhibition of cellgrowth. In certain embodiments, SRR G-1, or a derivative thereof, hasabout a 15 fold or greater increase in inhibition of cell growth. Incertain embodiments, SRR G-1, or a derivative thereof, has about a 20fold or greater increase in inhibition of cell growth. In certainembodiments, SRR G-1, or a derivative thereof, has about a 25 fold orgreater increase in inhibition of cell growth. In certain embodiments,SRR G-1, or a derivative thereof, has about a 30 fold or greaterincrease in inhibition of cell growth. In certain embodiments, SRR G-1,or a derivative thereof, has about a 35 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 40 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 45 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 50 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 55 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 60 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 65 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 70 fold or greater increase ininhibition of cell growth. In certain embodiments, SRR G-1, or aderivative thereof, has about a 75 fold or greater increase ininhibition of cell growth. Or a range between any two of these values.

In certain embodiments the compound of SRR G-1, or a derivative thereof,substantially free of its opposite enantiomer and at a concentration of500 nM has about a 39.5 fold or greater increase in inhibition of cellgrowth in a YUMM1.7 4 day growth assay as compared to the oppositeenantiomer of SRR G-1, or a derivative thereof.

In any of the embodiments described herein, SRR G-1, or a derivativethereof, possess greater desired pharmacological activity as compared toRSS G-1, or a derivative thereof. In any of the embodiments describedherein, the presence of RSS G-1, or a derivative thereof, would add tothe undesired pharmacological activity of a combination therapy with SRRG-1.

Pharmaceutical Compositions

Some embodiments herein are directed to a pharmaceutical or cosmeticcomposition comprising an enantiomerically purified SRR G-1, or aderivative thereof, of embodiments herein and a pharmaceutically orcosmetically acceptable carrier, adjuvant, or vehicle.

In some embodiments, the pharmaceutical or cosmetic compositionscomprising an enantiomerically purified SRR G-1, or a derivativethereof, for use in accordance with embodiments herein can be formulatedin conventional manner using one or more pharmaceutical or cosmeticallyacceptable carriers or excipients.

The carrier(s) must be “acceptable” in the sense of being compatiblewith the other ingredients of the formulation and not deleterious to therecipient thereof. Proper formulation is dependent upon the route ofadministration chosen. Any of the well-known techniques, carriers, andexcipients may be used as suitable and as understood in the art. Thepharmaceutical or cosmetic compositions comprising an enantiomericallypurified SRR G-1, or a derivative thereof, disclosed herein may bemanufactured in any manner known in the art, e.g., by means ofconventional mixing, dissolving, suspending, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or compressionprocesses.

Pharmaceutical or cosmetic compositions comprising an enantiomericallypurified SRR G-1, or a derivative thereof, include those suitable fororal, rectal, nasal, topical (including dermal, buccal, sublingual andintraocular), vaginal or parenteral (including intramuscular,sub-cutaneous and intravenous) administration. Compositions according tothe present invention may also be presented as a bolus, electuary orpaste. Tablets and capsules for oral administration may containconventional excipients such as binding agents, fillers, lubricants,disintegrants, or wetting agents. The tablets may be coated according tomethods well known in the art. Oral liquid preparations may be in theform of, for example, aqueous or oily suspensions, solutions, emulsions,syrups or elixirs, or may be presented as a dry product for constitutionwith water or other suitable vehicle before use. Such liquidpreparations may contain conventional additives such as suspendingagents, emulsifying agents, non-aqueous vehicles (which may includeedible oils), or preservatives. When desired, the above describedformulations may be adapted to provide sustained release characteristicsof the active ingredient(s) in the composition using standard methodswell-known in the art.

In the pharmaceutical composition comprising an enantiomericallypurified SRR G-1, or a derivative thereof, embodiments of the presentinvention, the compound(s) according to the present invention isformulated preferably in admixture with a pharmaceutically acceptablecarrier. In general, it is preferable to administer the pharmaceuticalcomposition comprising an enantiomerically purified SRR G-1, or aderivative thereof, orally, but certain pharmaceutical compositionscomprising an enantiomerically purified SRR G-1, or a derivativethereof, may be preferably administered parenterally and in particular,in intravenous or intramuscular dosage form, as well as via otherparenteral routes, such as transdermal, buccal, subcutaneous,suppository or other route, including via inhalation or intranasally.Oral dosage forms are preferably administered in tablet or capsule(preferably, hard or soft gelatin or other protein or polymer capsule)form. Intravenous and intramuscular pharmaceutical compositionscomprising an enantiomerically purified SRR G-1, or a derivativethereof, are preferably administered in sterile saline. Of course, oneof ordinary skill in the art may modify the formulations within theteachings of the specification to provide numerous pharmaceuticalcompositions comprising an enantiomerically purified SRR G-1, or aderivative thereof, for a particular route of administration withoutrendering the compositions of the present invention unstable orcompromising their therapeutic activity.

Pharmaceutical compositions comprising an enantiomerically purified SRRG-1, or a derivative thereof, suitable for parenteral injection maycomprise physiologically acceptable sterile aqueous or nonaqueoussolutions, dispersions, suspensions, or emulsions, or may comprisesterile powders for reconstitution into sterile injectable solutions ordispersions. Examples of suitable aqueous and nonaqueous carriers,diluents, solvents, or vehicles include water, ethanol, polyols(propylene glycol, polyethylene glycol, glycerol, and the like),suitable mixtures thereof, triglycerides, including vegetable oils suchas olive oil, or injectable organic esters such as ethyl oleate. Properfluidity can be maintained, for example, by the use of a coating such aslecithin, by the maintenance of the required particle size in the caseof dispersions, and/or by the use of surfactants.

These pharmaceutical or cosmetic compositions comprising anenantiomerically purified SRR G-1, or a derivative thereof, may alsocontain adjuvants such as preserving, wetting, emulsifying, and/ordispersing agents. Prevention of microorganism contamination of thecompositions can be accomplished by the addition of variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, sorbic acid, and the like. It may also bedesirable to include isotonic agents, for example, sugars, sodiumchloride, and the like. Prolonged absorption of injectablepharmaceutical or cosmetic compositions comprising an enantiomericallypurified SRR G-1, or a derivative thereof, can be brought about by theuse of agents capable of delaying absorption, for example, aluminummonostearate and/or gelatin.

Solid dosage forms of pharmaceutical compositions comprising anenantiomerically purified SRR G-1, or a derivative thereof, for oraladministration include capsules, tablets, powders, granules,stabilization in a polymer glass, dissolution in a lipid type liquid,dissolution in a solidified liquid, and dissolution in aself-emulsifying lipid. In such solid dosage forms, the active compoundis admixed with at least one inert customary excipient (or carrier) suchas sodium citrate or dicalcium phosphate or (a) fillers or extenders, asfor example, starches, lactose, sucrose, mannitol, or silicic acid; (b)binders, as for example, carboxymethylcellulose, alginates, gelatin,polyvinylpyrrolidone, sucrose, or acacia; (c) humectants, as forexample, glycerol; (d) disintegrating agents, as for example, agar-agar,calcium carbonate, potato or tapioca starch, alginic acid, certaincomplex silicates, or sodium carbonate; (e) solution retarders, as forexample, paraffin; (f) absorption accelerators, as for example,quaternary ammonium compounds; (g) wetting agents, as for example, cetylalcohol or glycerol monostearate; (h) adsorbents, as for example, kaolinor bentonite; and/or (i) lubricants, as for example, talc, calciumstearate, magnesium stearate, solid polyethylene glycols, sodium laurylsulfate, or mixtures thereof. In the case of capsules and tablets, thedosage forms may also comprise buffering agents.

Solid pharmaceutical compositions comprising an enantiomericallypurified SRR G-1, or a derivative thereof, of a similar type may also beused as fillers in soft or hard filled gelatin capsules using suchexcipients as lactose or milk sugar, as well as high molecular weightpolyethylene glycols, and the like.

Solid dosage forms of pharmaceutical compositions comprising anenantiomerically purified SRR G-1, or a derivative thereof, such astablets, dragees, capsules, and granules can be prepared with coatingsor shells, such as enteric coatings and others well known in the art.They may also contain opacifying agents, and can also be of suchcomposition that they release the active compound or compounds in adelayed manner. Examples of embedding compositions that can be used arepolymeric substances and waxes. The active compounds can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-mentioned excipients.

Liquid dosage forms of pharmaceutical compositions comprising anenantiomerically purified SRR G-1, or a derivative thereof, for oraladministration include pharmaceutically acceptable emulsions, solutions,suspensions, syrups, and elixirs. In addition to the active compounds,the liquid dosage form may contain inert diluents commonly used in theart, such as water or other solvents, solubilizing agents andemulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils, in particular,cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil,sesame seed oil, glycerol, tetrahydrofurfuryl alcohol, polyethyleneglycols, fatty acid esters of sorbitan, or mixtures of these substances,and the like.

Besides such inert diluents, the composition can also include adjuvants,such as wetting agents, emulsifying and suspending agents, sweetening,flavoring, and perfuming agents.

Suspensions, in addition to the active compound, may contain suspendingagents, as for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol or sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar, or tragacanth, or mixtures of thesesubstances, and the like.

Pharmaceutical compositions comprising an enantiomerically purified SRRG-1, or a derivative thereof, for rectal or vaginal administration,where applicable, can be prepared by mixing an active agent and anyadditional compounds with suitable non-irritating excipients or carrierssuch as cocoa butter, polyethylene glycol or a suppository wax, whichare solid at ordinary room temperature, but liquid at body temperature,and therefore, melt in the rectum or vaginal cavity and release theactive.

Dosage forms of pharmaceutical or cosmetic compositions comprising anenantiomerically purified SRR G-1, or a derivative thereof, for topicaladministration include ointments, powders, sprays inhalants, and dropssuitable for administration to the eye, ear or nose. The compound(s) areadmixed under sterile conditions with a physiologically acceptablecarrier, and any preservatives, buffers, and/or propellants that may berequired. Opthalmic formulations, eye ointments, powders, and solutionsare also contemplated as being within the scope of this invention.

In some embodiments, the pharmaceutical or cosmetic compositionscomprising an enantiomerically purified SRR G-1, or a derivativethereof, for use in accordance with embodiments herein additionallyinclude at least one sun-blocking agent. In certain embodiments, thepharmaceutical composition further comprises at least sunscreen lotion.In oilier embodiments, the pharmaceutical or cosmetic compositioncomprises a formulated sunblock or sunscreen lotion and enantiomericallypurified SRR G-1, or a derivative thereof.

The active compound or pharmaceutical or cosmetic composition comprisingan enantiomerically purified SRR G-1, or a derivative thereof, can beeffective over a wide dosage range and can be generally administered ina therapeutically effective amount. It will be understood, however, thatthe amount of the compound or composition actually administered willusually be determined by a physician, according to the relevantcircumstances, including the condition to be treated, the chosen routeof administration, the actual compound or composition administered, theage, weight, and response of the individual patient, the severity of thepatient's symptoms, and the like.

In some embodiments, the pharmaceutical or cosmetic compositioncomprising an enantiomerically purified SRR G-1, or a derivativethereof, may comprise about 0.001% to about 50% of one or more compoundsor compositions disclosed herein. In some embodiments, the one or morecompounds or compositions is in an amount of about 0.001% to about 50%,about 0.001% to about 45%, about 0.001% to about 40%, about 0.001% toabout 30%, about 0.001% to about 20%, about 0.001% to about 10%, about0.001% to about 5%, about 0.01% to about 50%, about 0.01% to about 45%,about 0.01% to about 40%, about 0.01% to about 30%, about 0.01% to about20%, about 0.01% to about 10%, about 0.01% to about 5%, about 0.05% toabout 50%, about 0.05% to about 45%, about 0.05% to about 40%, about0.05% to about 30%, about 0.05% to about 20%, about 0.05% to about 10%,about 0.1% to about 50%, about 0.1% to about 45%, about 0.1% to about40%, about 0.1% to about 30%, about 0.1% to about 20%, about 0.1% toabout 10%, about 0.1% to about 5%, about 0.5% to about 50%, about 0.5%to about 45%, about 0.5% to about 40%, about 0.5% to about 30%, about0.5% to about 20%, about 0.5% to about 10%, about 0.5% to about 5%,about 1% to about 50%, about 1% to about 45%, about 1% to about 40%,about 1% to about 35%, about 1% to about 30%, about 1% to about 25%,about 1% to about 20%, about 1% to about 15%, about 1% to about 10%,about 1% to about 5%, about 5% to about 45%, about 5% to about 40%,about 5% to about 35%, about 5% to about 30%, about 5% to about 25%,about 5% to about 20%, about 5% to about 15%, about 5% to about 10%,about 10% to about 45%, about 10% to about 40%, about 10% to about 35%,about 10% to about 30%, about 10% to about 25%, about 10% to about 20%,about 10% to about 15%, or a value within one of these ranges. Specificexamples may include about 0.001%, about 0.01%, about 0.05%, about 0.1%,about 0.25%, about 0.5%, about 0.75%, about 1%, about 5%, about 10%,about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about45%, about 50%, about 60%, about 70%, about 80%, about 90%, or a rangebetween any two of these values. The foregoing all representing weightpercentages of the composition. In some embodiments, the composition issuitable for topical administration. In some embodiments, thecomposition is suitable for oral administration. In some embodiments,the composition is suitable for oral, parenteral (includingsubcutaneous, intradermal, intramuscular, intravenous, intraarticular,and intramedullary), intraperitoneal, transmucosal, transdermal, rectal,intranasal, topical (including dermal, buccal, sublingual andintraocular), or intravaginal administration.

In some embodiments, the compound or pharmaceutical or cosmeticcompositions comprising an enantiomerically purified SRR G-1, or aderivative thereof, is in a therapeutically effective amount. In someembodiments, the therapeutically effective amount may be about 0.01 mgto about 1000 mg, about 0.01 mg to about 900 mg, about 0.01 mg to about800 mg, about 0.01 mg to about 700 mg, about 0.01 mg to about 600 mg,about 0.01 mg to about 500 mg, about 0.01 mg to about 400 mg, about 0.01mg to about 300 mg, about 0.01 mg to about 200 mg, about 0.01 mg toabout 100 mg, 0.1 mg to about 1000 mg, about 0.1 mg to about 900 mg,about 0.1 mg to about 800 mg, about 0.1 mg to about 700 mg, about 0.1 mgto about 600 mg, about 0.1 mg to about 500 mg, about 0.1 mg to about 400mg, about 0.1 mg to about 300 mg, about 0.1 mg to about 200 mg, about0.1 mg to about 100 mg, about 1 mg to about 1000 mg, about 1 mg to about900 mg, about 1 mg to about 800 mg, about 1 mg to about 700 mg, about 1mg to about 600 mg, about 1 mg to about 500 mg, about 1 mg to about 400mg, about 1 mg to about 300 mg, about 1 mg to about 200 mg, about 1 mgto about 100 mg, about 10 mg to about 1000 mg, about 50 mg to about 1000mg, about 100 mg to about 1000 mg, about 200 mg to about 1000 mg, about300 mg to about 1000 mg, about 400 mg to about 1000 mg, about 500 mg toabout 1000 mg, about 10 mg to about 500 mg, about 50 mg to about 500 mg,about 100 mg to about 500 mg, about 10 mg to about 300 mg, about 50 mgto about 300 mg, from about 100 mg to about 300 mg, about 10 mg to about150 mg, about 50 mg to about 150 mg, about 60 mg to about 120 mg, about50 mg to about 120 mg or a range between any two of these values.Specific examples include, for example, about 1000 mg, about 900 mg,about 800 mg, about 700 mg, about 750 mg, about 600 mg, about 500 mg,about 400 mg, about 450 mg, about 300 mg, about 250 mg, about 200 mg,about 175 mg, about 150 mg, about 125 mg, about 120 mg, about 110 mg,about 100 mg, about 90 mg, about 80 mg, about 70 mg, about 60 mg, about50 mg, about 30 mg, about 20 mg, about 10 mg, about 5 mg, about 1 mg,about 0.1 mg, about 0.01 mg, or any value between the ranges disclosedabove.

In some embodiments, the therapeutically effective amount can varyaccording to, for example, the particular use for which the treatment ismade, the manner of administration of the compound or composition, thehealth and condition of the patient, and the judgment of the prescribingphysician. The proportion or concentration of a compound or compositionin a pharmaceutical or cosmetic composition comprising anenantiomerically purified SRR G-1, or a derivative thereof, can varydepending upon a number of factors including dosage, chemicalcharacteristics (e.g., hydrophobicity), and the route of administration.For example, the compounds or compositions can be provided in an aqueousphysiological buffer solution containing about 0.1 to about 10% w/v ofthe compound or composition for parenteral administration. Some typicaldose ranges for the compounds or compositions are from about 1 μg/kg toabout 1 g/kg of body weight per day. In some embodiments, the dose rangeis from about 0.01 mg/kg to about 100 mg/kg of body weight per day. Thedosage is likely to depend on such variables as the type and extent ofprogression of the disease or disorder, the overall health status of theparticular patient, the relative biological efficacy of the compound orcomposition selected, formulation of the excipient, and its route ofadministration. Effective doses can be extrapolated from dose-responsecurves derived from in vitro or animal model test systems.

The amount of compound or pharmaceutical or cosmetic compositioncomprising an enantiomerically purified SRR G-1, or a derivativethereof, administered to a patient will vary depending upon what isbeing administered, the purpose of the administration, such asprophylaxis or therapy, the state of the patient, the manner ofadministration, and the like. In therapeutic applications, compositionscan be administered to a patient already suffering from a disease in anamount sufficient to cure or at least partially arrest the symptoms ofthe disease and its complications.

In any of the pharmaceutical compositions comprising an enantiomericallypurified SRR G-1, or a derivative thereof, described herein may have oneor more additional therapeutic agents.

The additional therapeutic agents may be selected from the group, but isnot limited to, consisting of a weight loss drug, an antihyperglycemicdrug, an insulin sensitizer, a glucagon-like peptide 1 (GLP1) receptoragonist, a sodium glucose cotransporter 2 (SGLT2) inhibitor, insulin, aninsulin analogue, sulfonylureas, a dipeptidyl peptidase 4 (DPP4)inhibitor, an alphaglucosidase inhibitor (AGI), a bile acid sequestrant(BAS), sympatholytic dopamine receptor agonist, incretins, ahypertension drug, a lipid-modifying agent, an anti-obesity agent, animmunotherapy agent, a chemotherapy agent, a targeted kinase inhibitor,a histone deacetylase inhibitor, an anti-infective agent, a bromodomaininhibitor, and combinations thereof.

The immunotherapy agent may be selected from the group, but is notlimited to, consisting of PD-1 inhibitors (Pembrolizumab, Nivolumab,anti-PD-1), PD-L1 inhibitors (i.e. Atezolizumab, Avelumab, Durvalumab,anti-PD-L1), CTLA-4 inhibitors (i.e. Ipilimumab, anti-B7-1/B7-2,anti-CTLA-4), IL-2, IL-7, IL-12, Oncolytic Viruses (TalimogeneLaherparepvec), cytosine phosphate-guanosine, oligodeoxynucleotides,Imiquimod, Resiquimod, and antibodies targeting T cell immunoreceptorwith Ig and ITIM domains (TIGIT), inducible co-stimulator (ICOS),Lymphocyte activation gene 3 (LAG-3), T-cell immunoglobulin and Mucindomain containing molecule 3 (TIM3), V-domain containing IG supressor ofT cella ctivation (VISTA), OX40, Glucocorticoid-induced TNF receptor(GITR), CD40, CD47, CD94/NKG2A, Killer immunoglobulin receptor (KIR),and combinations thereof.

The chemotherapy agent may be selected from the group, but is notlimited to, consisting of Cyclophosphamide, methotrexate,5-fluorouracil, Doxorubicin, Docetaxel, bleomycin, vinblastine,dacarbazine, Mustine, vincristine, procarbazine, etoposide, cisplatin,Epirubicin, capecitabine, folinic acid, oxaliplatin, temozolomide,taxanes, and combinations thereof.

The targeted kinase inhibitor may be selected from the group, but is notlimited to, consisting of Vemurafenib, Dabrafenib, Trametinib,Vandetanib, SU6656, Sunitinib, Sorafenib, Selumetinib, Ruxolitinib,Pegaptanib, Pazopanib, Nilotinib, Mubritinib, Lenvatinib, Lapatinib,Imatinib, Ibrutinib, Gefitinib, Fostamatinib, Erlotinib, Erdafitinib,Dasatinib, Cabozantinib, Crizotinib, Cobimetinib, Cetuximab, Bosutinib,Binimetinib, Axitinib, Afatinib, Adavosertib, and combinations thereof.

The histone deacetylase inhibitor may be selected from the group, but isnot limited to, consisting of Vorinostat, Romidepsin, Chidamide,Panobinostat, Belinostat, Valproic acid, Givinostat, and combinationsthereof.

The anti-infective agent may be selected from the group, but is notlimited to, consisting of oritavancin (Orbactiv), dalvavancin(Dalvance), tedizolid phosphate, (Sivextro), clindamycin, linezolid(Zyvox), mupirocin (Bactroban), trimethoprim, sulfamethoxazole,trimethoprim-sulfamethoxazole (Septra or Bactrim), a tetracycline,vancomycin, daptomycin, fluoroquinolines, and combinations thereof.

The bromodomain inhibitor may be selected from the group, but is notlimited to, consisting of OTX015/MK-8628, CPI-0610, BMS-986158,ZEN003694, GSK2820151, GSK525762, INCB054329, INCB057643, ODM-207,R06870810, BAY1238097, CC-90010, AZD5153, FT-1101, ABBV-744, RVX-000222,and combinations thereof.

Methods of Use

Provided herein is a method of treating or preventing a disease ordisorder in a subject in need thereof comprising administering to asubject a therapeutically effective amount of a compound orpharmaceutical composition comprising enantiomerically purified SRR G-1,or a derivative thereof, according to any embodiment disclosed herein.

Methods of treating or preventing a disease or disorder in a subject inneed thereof comprising administering to a subject a therapeuticallyeffective amount of a compound or pharmaceutical composition comprisingenantiomerically purified SRR G-1, or a derivative thereof, describedherein where treatment with SRR G-1 is acting as an adjuvant prior to,with, or after one or more additional therapies selected from surgicaltherapy, chemotherapy, anti-PD-1 therapy, targeted molecular oranti-proliferative therapy or radiofrequency ablation therapy.

Some embodiments describe a method wherein the cancer or cells causingor involved in the disease or disorder expresses GPER.

In any embodiments described herein the subject is a human or an animal.

In some embodiments, said disease or disorder is selected from the groupconsisting of cancer, endometritis, prostatitis, polycystic ovariansyndrome, urinary incontinence, hormone-related disorders, hearingdisorders, hot flashes, profuse sweating, hypertension, stroke,ischemia, myocardial infarction, dilated cardiomyopathy, obesity,insulin resistance, osteoporosis, atherosclerosis, symptoms ofmenopause, inflammation, rheumatoid arthritis, osteoarthritis,lymphoproliferative disorders, myeloproliferative disorders,eosinophilia, histiocytosis, paroxysmal nocturnal hemoglobinuria,systemic mastocytosis, venous thrombosis, embolisms, depression,insomnia, anxiety, neuropathy, multiple sclerosis, Parkinson's disease,Alzheimer's disease, inflammatory bowel disease, Crohn's disease, celiacdisease, proteinuric renal disease, vascular disease, and thymicatrophy.

Some embodiments describe a method of preventing or reducing thelikelihood of pregnancy after intercourse comprising administering to asubject a therapeutically effective amount of a compound or composition,or a derivative thereof, according to any embodiment disclosed herein.

Some embodiments describe a method of restoring the lipid profile in asubject in need thereof comprising administering to a subject atherapeutically effective amount of a compound or composition, or aderivative thereof, according to any embodiment disclosed herein.

Some embodiments describe a method of treating or preventing type 2diabetes in a subject in need thereof comprising administering to asubject a therapeutically effective amount of a compound or composition,or a derivative thereof, according to any embodiment disclosed herein.

Type 2 diabetes is a disease diagnosed by a set of characteristicsselected from the group consisting of an A1C level of greater than orequal to 6.5%, a fasting plasma glucose (FPG) amount of greater than 126mg/dL, and an oral glucose tolerance test (OGTT) amount of greater than200 mg/dL. Subjects with type 2 diabetes are at higher risk ofdeveloping dyslipidemia, hypertension, and artheroscleroticcardiovascular disease (ASCVD). In embodiments, the subject is treatedby the administration of compound or composition, or a derivativethereof, according to any embodiment disclosed herein wherein thesymptoms of diabetes is treated. In embodiments, the subject is treatedby the administration of a compound or composition, or a derivativethereof, according to any embodiment disclosed herein wherein the A1Clevel is reduced to less than 6.5%, between 6.4% and 5.7%, or less than5.7%. In embodiments, the subject is treated by the administration of acompound or composition, or a derivative thereof, according to anyembodiment disclosed herein wherein the fasting plasma glucose (FPG) isreduced to less than 126 mg/dL, between 125 mg/dL to 110 mg/dL, lessthan 110 mg/dL, or less than 100 mg/dL. In embodiments, the subject istreated by the administration of a compound or composition, or aderivative thereof, according to any embodiment disclosed herein whereinthe oral glucose tolerance test (OGTT) is reduced to less than 200mg/dL, between 199 mg/dL and 140 mg/dL, or less than 140 mg/dL. Inembodiments, the subject is treated by the administration of a compoundor composition, or a derivative thereof, according to any embodimentdisclosed herein wherein the blood pressure is reduced to less than130/80 mmHg, less than 120/80 mmHg, less than 110/80 mmHg, or less than100/80 mmHg. In embodiments, the subject is treated by theadministration of a compound or composition, or a derivative thereof,according to any embodiment disclosed herein wherein the blood glucoselevel is reduced to less than 70 mg/dL, or less than 50 mg/dL. Inembodiments, the subject is treated by the administration of a compoundor composition, or a derivative thereof, according to any embodimentdisclosed herein wherein the risk of developing dyslipidemia,hypertension, or artherosclerotic cardiovascular disease (ASCVD) isreduced by about 5%, about 10%, about 15%, about 20%, about 25%, about30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%,about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about95%, or about 100%.

Pre-diabetes is diagnosed based upon a set of characteristics selectedfrom the group consisting of an A1C level of about 5.7% to about 6.4%, afasting plasma glucose (FPG) amount of about 100 mg/dL to about 125mg/dL, and an oral glucose tolerance test (OGTT) amount of about 140mg/dL to about 200 mg/dL. A pre-diabetic subject can also be diagnosedwith impaired glucose tolerance, impaired fasting glucose, or insulinresistance. Subjects with pre-diabetes are at higher risk of developinghyperglycemia, dyslipidemia, hypertension, artheroscleroticcardiovascular disease (ASCVD), cardiometabolic disease, chronic kidneydisease, early nephropathy, retinopathy, cardiovascular disease andbiomechanical complications. In embodiments, the subject is treated bythe administration of a compound or composition, or a derivativethereof, according to any embodiment disclosed herein wherein thesymptoms of pre-diabetes is treated. In embodiments, the subject istreated by the administration of a compound or composition, or aderivative thereof, according to any embodiment disclosed herein whereinthe A1C level is reduced to less than 6.4%, or less than 5.7%. Inembodiments, the subject is treated by the administration of a compoundor composition, or a derivative thereof, according to any embodimentdisclosed herein wherein the fasting plasma glucose (FPG) is reduced toless than 125 mg/dL, less than 110 mg/dL, or less than 100 mg/dL. Inembodiments, the subject is treated by the administration of a compoundor composition, or a derivative thereof, according to any embodimentdisclosed herein wherein the oral glucose tolerance test (OGTT) isreduced to less than 199 mg/dL, or less than 140 mg/dL. In embodiments,the subject is treated by the administration of a compound orcomposition, or a derivative thereof, according to any embodimentdisclosed herein wherein the blood pressure is reduced to less than130/80 mmHg, less than 120/80 mmHg, less than 110/80 mmHg, or less than100/80 mmHg. In embodiments, the subject is treated by theadministration of a compound or composition, or a derivative thereof,according to any embodiment disclosed herein wherein the blood glucoselevel is reduced to less than 70 mg/dL, or less than 50 mg/dL. Inembodiments, the subject is treated by the administration of a compoundor composition, or a derivative thereof, according to any embodimentdisclosed herein wherein the risk of developing hyperglycemia,dyslipidemia, hypertension, artherosclerotic cardiovascular disease(ASCVD), cardiometabolic disease, chronic kidney disease, earlynephropathy, retinopathy, cardiovascular disease or biomechanicalcomplications is reduced by about 5%, about 10%, about 15%, about 20%,about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%,about 90%, about 95%, or about 100%.

Conditions characterized by an increase in the levels of A1C, glucose,insulin, homeostasis model of assessment of insulin resistance(HOMA-IR), urinary 8-iso-PGF2α, oxidative stress in adipose tissue, andcarbonylation of GLUT4 lead to a diagnosis of impaired glucosetolerance, impaired fasting glucose, insulin resistance, pre-diabetes,or type 2 diabetes. Subjects with a condition as described herein are athigher risk of developing pre-diabetes, type 2 diabetes, hyperglycemia,dyslipidemia, hypertension, artherosclerotic cardiovascular disease(ASCVD), cardiometabolic disease, chronic kidney disease, earlynephropathy, retinopathy, cardiovascular disease and biomechanicalcomplications. In embodiments, the subject is treated by theadministration of a compound or composition, or a derivative thereof,according to any embodiment disclosed herein wherein the symptoms of thecondition is treated. In embodiments, the subject is treated by theadministration of a compound or composition, or a derivative thereof,according to any embodiment disclosed herein wherein the A1C level isreduced to less than 6.4%, or less than 5.7%. In embodiments, thesubject is treated by the administration of a compound or composition,or a derivative thereof, according to any embodiment disclosed hereinwherein the fasting plasma glucose (FPG) is reduced to less than 125mg/dL, less than 110 mg/dL, or less than 100 mg/dL. In embodiments, thesubject is treated by the administration of a compound or composition,or a derivative thereof, according to any embodiment disclosed hereinwherein the oral glucose tolerance test (OGTT) is reduced to less than199 mg/dL, or less than 140 mg/dL. In embodiments, the subject istreated by the administration of a compound or composition, or aderivative thereof, according to any embodiment disclosed herein whereinthe blood pressure is reduced to less than 130/80 mmHg, less than 120/80mmHg, less than 110/80 mmHg, or less than 100/80 mmHg. In embodiments,the subject is treated by the administration of a compound orcomposition, or a derivative thereof, according to any embodimentdisclosed herein wherein the blood glucose level is reduced to less than70 mg/dL, or less than 50 mg/dL. In embodiments, the subject is treatedby the administration of a compound or composition, or a derivativethereof, according to any embodiment disclosed herein wherein the riskof developing pre-diabetes, type 2 diabetes, hyperglycemia,dyslipidemia, hypertension, artherosclerotic cardiovascular disease(ASCVD), cardiometabolic disease, chronic kidney disease, earlynephropathy, retinopathy, cardiovascular disease and biomechanicalcomplications is reduced by about 5%, about 10%, about 15%, about 20%,about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%,about 90%, about 95%, or about 100%.

In embodiments, the one or more additional therapeutic agents may beselected from the group consisting of a weight loss drug, anantihyperglycemic drug, an insulin sensitizer, a glucagon-like peptide 1(GLP1) receptor agonist, a sodium glucose cotransporter 2 (SGLT2)inhibitor, insulin, an insulin analogue, sulfonylureas, a dipeptidylpeptidase 4 (DPP4) inhibitor, an alphaglucosidase inhibitor (AGI), abile acid sequestrant (BAS), sympatholytic dopamine receptor agonist,incretins, a hypertension drug, a lipid-modifying agent, andcombinations thereof.

In embodiments, the weight loss drug is selected from the groupconsisting of diethyproprion, phendimetrazine, phentermine, orlistat,phentermine/topiramate extended release (ER), lorcaserin, naltrexoneER/bupropion ER, and liraglutide. In embodiments, diethyproprion isadministered at 25 mg. In embodiments, phendimetrazine is administeredat 35 mg or 105 mg. In embodiments, phentermine is administered at 8 mg,15 mg, 30 mg, or 37.5 mg. In embodiments, orlistat is administered at 60mg or 120 mg. In embodiments, phentermine/topiramate extended release isadministered at phentermine 3.75 mg/topiramate 23 mg, phentermine 7.5mg/topiramate 46 mg daily, or phentermine 15 mg/topiramate 92 mg. Inembodiments, lorascerin is administered at 10 mg or 20 mg. Inembodiments, naltrexone ER/bupropion ER is administered at 8 mgnaltrexone/90 mg bupropion. In embodiments, liraglutide is administeredat 1.2 mg, 1.8 mg, or 3 mg.

In embodiments, the antihyperglycemic drug is selected from the groupconsisting of metformin and acarbose. In embodiments, metformin isadministered at 500 mg, 625 mg, 750 mg, 850 mg, 2000 mg, 2500 mg, or 1gram. In embodiments, acarbose is administered at 25 mg, 50 mg, or 100mg.

In embodiments, the insulin sensitizer is selected from the groupconsisting of thiazolidinediones (TZDs), pioglitazone, androsiglitazone. In embodiments, pioglitazone is administered at 15 mg, 30mg, or 45 mg. In embodiments, rosiglitazone is administered at 2 mg, 4mg, or 8 mg.

In embodiments, the glucagon-like peptide 1 (GLP1) receptor agonist isselected from the group consisting of liraglutide, exenatide,albiglutide, and dulaglutide. In embodiments, liraglutide isadministered at 1.2 mg, 1.8 mg, or 3 mg. In embodiments, exenatide isadministered at 2 mg. In embodiments, albiglutide is administered at 30mg or 50 mg. In embodiments, dulaglutide is administered at 0.75 mg or1.5 mg.

In embodiments, the sodium glucose cotransporter 2 (SGLT2) inhibitor isselected from the group consisting of empagliflozin, canagliflozin, anddapagliflozin. In embodiments, empagliflozin is administered at 5 mg, 10mg, 12.5 mg, or 25 mg. In embodiments, canagliflozin is administered at50 mg, 100 mg, 150 mg, or 300 mg.

In embodiments, the insulin is selected from the group consisting ofinsulin analogues, basal insulin analogues, neutral protamine Hagedorn(NPH), rapid acting insulin analogues, and inhaled insulin.

In embodiments, the insulin analogue is selected from the groupconsisting of glargine, degludec, and detemir. In embodiments, glargineis administered at 100 units or 300 units. In embodiments, degludec isadministered at 30 units, 100 units, 200 units, 300 units, or 600 units.In embodiments, detemir is administered at 100 units or 300 units.

In embodiments, the rapid acting insulin analogue is selected from thegroup consisting of lispro, aspart, and glulisine. In embodiments,lispro is administered at 50 units, 75 units, 100 units, 300 units, or1000 units. In embodiments, aspart is administered at 50 units, 90units, 210 units, 300 units, 700 units, or 1000 units. In embodiments,glulisine is administered at 300 units or 1000 units.

In embodiments, the sulfonylureas is selected from the group consistingof acetohexamide, carbutamide, chlorpropamide, glycyclamide(tolhexamide), metahexamide, tolazamide, tolbutamide, glibenclamide(glyburide), glibornuride, gliclazide, glipizide, gliquidone,glisoxepide, glyclopyramide, and glimepiride. In embodiments,acetohexamide is administered at 250 mg or 500 mg. In embodiments,carbutamide is administered at 250 mg or 500 mg. In embodiments,chlorpropamide is administered at 100 mg or 250 mg. In embodiments,tolazamide is administered at 100 mg, 250 mg, or 500 mg. In embodiments,tolbutamide is administered at 250 mg or 500 mg. In embodiments,glibenclamide is administered at 5 mg. In embodiments, glipizide isadministered at 2.5 mg, 5 mg, or 10 mg. In embodiments, glimepiride isadministered at 1 mg, 2 mg, 3 mg, 4 mg, 6 mg, or 8 mg.

In embodiments, the dipeptidyl peptidase 4 (DPP4) inhibitor is selectedfrom the group consisting of linagliptin, saxagliptin, and alogliptin.In embodiments, linagliptin is administered at 2.5 mg, 5 mg, 10 mg, or25 mg. In embodiments, saxagliptin is administered at 2.5 mg or 5 mg. Inembodiments, alogliptin is administered at 6.25 mg, 12.5 mg, or 25 mg.

In embodiments, the alpha glucosidase inhibitor (AGI) is selected fromthe group consisting of acarbose, miglitol, and voglibose. Inembodiments, acarbose is administered at 25 mg, 50 mg, or 100 mg. Inembodiments, miglitol is administered at 25 mg, 50 mg, or 100 mg. inembodiments, voglibose is administered at 0.2 mg or 0.3 mg.

In embodiments, the bile acid sequestrant (BAS) is selected from thegroup consisting of cholestyramine, colestipol, and colesevelam. Inembodiments, cholestyramine is administered at 800 mg, 1 gram, or 4grams. In embodiments, colestipol is administered at 1 gram or 5 grams.In embodiments, colesevelam is administered at 375 mg, 625 mg, 1.875grams, or 3.75 grams.

In embodiments, the sympatholytic dopamine receptor agonist isbromocriptine mesylate. In embodiments, bromocriptine mesylate isadministered at 0.8 mg, 2.5 mg, or 5 mg.

In embodiments, the hypertension drug is selected from the groupconsisting of angiotensin-converting enzyme inhibitors (ACEIs),angiotensin II receptor blockers (ARBs), beta blockers, calcium channelblockers (CCBs), and thiazide diuretics.

In embodiments, the lipid-modifying agent is selected from the groupconsisting of ezetimibe, simvastatin, monoclonal antibody inhibitors ofproprotein convertase subtilisin-kexin type 9 serine protease (PCSK9),evolocumab, alirocumab, fibrates, niacin, eicosapentaenoic acid (EPA),docosahexaenoic acid (DHA) and omega-3 fatty acids. In embodiments,ezetimibe is administered at 10 mg. In embodiments, simvastatin isadministered at 5 mg, 10 mg, 20 mg, 40 mg, or 80 mg. In embodiments,evolocumab is administered at 140 mg or 420 mg. In embodiments,alirocumab is administered at 75 mg, 150 mg, or 300 mg. In embodiments,niacin is administered at 375 mg, 500 mg, 750 mg, or 1 gram.

Treatment efficacy may be assessed by measuring the level of insulin inthe blood. A normal fasting insulin level is below 5. A fasting insulinlevel around 8.0 results in twice the risk of pre-diabetes, and afasting insulin of about 25 results in about a five times the risk ofprediabetes. In embodiments, administration of a compound orcomposition, or a derivative thereof, according to any embodimentdisclosed herein to a subject in need thereof decreases the fastinginsulin level is less than 5, less than 8, or less than 25.

Urinary 8-iso-PGF2a is a well-established marker of oxidativestress-induced lipid peroxidation. A rise in urinary 8-iso-PGF2aindicates the development of systemic oxidative stress. Treatmentefficacy may be assessed by measuring the level of urinary 8-iso-PGF2αin adipose tissue. In embodiments, administration of a compound orcomposition, or a derivative thereof, according to any embodimentdisclosed herein to a subject in need thereof decreases the level ofurinary 8-iso-PGF2α.

Treatment efficacy may be assessed by measuring the level of oxidativestress in adipose tissue. Oxidative stress is measured by an increase inany one of the following enzymes: superoxide dismutase 2 (SOD2),catalase, glutathione peroxidase, peroxiredoxin, aldehyde dehydrogenase,aldo-keto reductase, and glutathione S-transferase. In embodiments,administration of a compound or composition, or a derivative thereof,according to any embodiment disclosed herein to a subject in needthereof decreases the level of one or more of the following enzymes:superoxide dismutase 2 (SOD2), catalase, glutathione peroxidase,peroxiredoxin, aldehyde dehydrogenase, aldo-keto reductase, andglutathione S-transferase.

Treatment efficacy may be assessed by measuring the level ofcarbonylation of GLUT4. In adipose tissue during overnutrition,oxidative stress results in extensive oxidation and carbonylation ofnumerous proteins, including carbonylation of GLUT4 near the glucosetransport channel, which results in the loss of GLUT4 activity. Thecarbonylation and oxidation-induced inactivation of GLUT4 may result ininsulin resistance. In embodiments, administration of a compound orcomposition, or a derivative thereof, according to any embodimentdisclosed herein to a subject in need thereof decreases the level ofGLUT4 carbonylation.

Some embodiments describe a method of treating or preventing cancer,preventing the reoccurrence of cancer, inhibiting the progression ofcancer, shrinking a cancer prior to additional therapy, or reducingcirculating tumor cells or metastases prior to additional therapy in asubject in need thereof comprising administering to a subject atherapeutically effective amount of a compound or composition, or aderivative thereof, according to any embodiment disclosed herein.

In some embodiments, said cancer is selected from the group consistingof reproductive cancers, hormone-dependent cancers, leukemia, colorectalcancer, prostate cancer, breast cancer, ovarian carcinoma, endometrialcancer, uterine carcinosarcoma, stomach cancer, rectal cancer, livercancer, pancreatic cancer, lung cancer, uterine cancer, cervical cancer,cervix uteri cancer, corpus uteri cancer, ovary cancer, testicularcancer, bladder cancer, renal cancer, brain/CNS cancer, head and neckcancer, throat cancer, Hodgkin's disease, non-Hodgkin's lymphoma,multiple myeloma, melanoma, acute leukemia, lymphocytic leukemia, hairycell leukemia, acute myelogenous leukemia, Ewing's sarcoma, small celllung cancer, non-small cell lung cancer, choriocarcinoma,rhabdomyosarcoma, Wilms's Tumor, neuroblastoma, cancer of themouth/pharynx, cancer of the esophagus, cancer of the larynx, kidneycancer, lymphoma, Burkitt lymphoma, sarcoma, angiosarcoma, glioblastoma,medulloblastoma, astrocytoma, and Merkel cell carcinoma.

In particular embodiment, the cancer is selected from the groupconsisting of melanoma, colorectal cancer, non-small cell lung cancer,and pancreatic cancer.

Some embodiments describe a method increasing, or preventing orreversing loss of, skin pigmentation in a subject in need thereofcomprising administering to a subject a therapeutically effective amountof a compound or composition, or a derivative thereof, according to anyembodiment disclosed herein.

Some embodiments describe a method of skin protection in a subject inneed thereof comprising administering to a subject a therapeuticallyeffective amount of a compound or composition, or a derivative thereof,according to any embodiment disclosed herein.

Some embodiments describe a method of skin protection comprisingincreasing skin pigmentation in a subject in need thereof comprisingadministering to a subject a therapeutically effective amount of acompound or composition, or a derivative thereof, according to anyembodiment disclosed herein.

Some embodiments describe a method of protection of skin from skincancer in a subject in need thereof comprising administering to asubject a therapeutically effective amount of a compound or composition,or a derivative thereof, according to any embodiment disclosed herein.

Some embodiments describe a method of protection of skin from skincancer comprising increasing skin pigmentation in a subject in needthereof comprising administering to a subject a therapeuticallyeffective amount of a compound or composition, or a derivative thereof,according to any embodiment disclosed herein.

In embodiments, the methods may include the co-administration of one ormore additional therapeutic agents. In embodiments, co-administrationmay be part of the same pharmaceutical composition comprising anenantiomerically purified SRR G-1, or a derivative thereof, or separatepharmaceutical compositions comprising an enantiomerically purified SRRG-1, or a derivative thereof, described herein. In embodiments,co-administration may be at the same time, substantially the same time,before or after administration of the compositions described herein.

The additional therapeutic agents may be selected from the groupconsisting of a weight loss drug, an antihyperglycemic drug, an insulinsensitizer, a glucagon-like peptide 1 (GLP1) receptor agonist, a sodiumglucose cotransporter 2 (SGLT2) inhibitor, insulin, an insulin analogue,sulfonylureas, a dipeptidyl peptidase 4 (DPP4) inhibitor, analphaglucosidase inhibitor (AGI), a bile acid sequestrant (BAS),sympatholytic dopamine receptor agonist, incretins, a hypertension drug,a lipid-modifying agent, an anti-obesity agent, an immunotherapy agent,a chemotherapy agent, a targeted kinase inhibitor, a histone deacetylaseinhibitor, an anti-infective agent, a bromodomain inhibitor, andcombinations thereof.

The immunotherapy agent may be selected from the group consisting ofPD-1 inhibitors (Pembrolizumab, Nivolumab, anti-PD-1), PD-L1 inhibitors(i.e. Atezolizumab, Avelumab, Durvalumab, anti-PD-L1), CTLA-4 inhibitors(i.e. Ipilimumab, anti-B7-1/B7-2, anti-CTLA-4), IL-2, IL-7, IL-12,Oncolytic Viruses (Talimogene Laherparepvec), cytosinephosphate-guanosine, oligodeoxynucleotides, Imiquimod, Resiquimod, andantibodies targeting T cell immunoreceptor with Ig and ITIM domains(TIGIT), inducible co-stimulator (ICOS), Lymphocyte activation gene 3(LAG-3), T-cell immunoglobulin and Mucin domain containing molecule 3(TIM3), V-domain containing IG supressor of T cella ctivation (VISTA),OX40, Glucocorticoid-induced TNF receptor (GITR), CD40, CD47,CD94/NKG2A, Killer immunoglobulin receptor (KIR), and combinationsthereof.

The chemotherapy agent may be selected from the group consisting ofCyclophosphamide, methotrexate, 5-fluorouracil, Doxorubicin, Docetaxel,bleomycin, vinblastine, dacarbazine, Mustine, vincristine, procarbazine,etoposide, cisplatin, Epirubicin, capecitabine, folinic acid,oxaliplatin, temozolomide, taxanes, and combinations thereof.

The targeted kinase inhibitor may be selected from the group consistingof Vemurafenib, Dabrafenib, Trametinib, Vandetanib, SU6656, Sunitinib,Sorafenib, Selumetinib, Ruxolitinib, Pegaptanib, Pazopanib, Nilotinib,Mubritinib, Lenvatinib, Lapatinib, Imatinib, Ibrutinib, Gefitinib,Fostamatinib, Erlotinib, Erdafitinib, Dasatinib, Cabozantinib,Crizotinib, Cobimetinib, Cetuximab, Bosutinib, Binimetinib, Axitinib,Afatinib, Adavosertib, and combinations thereof.

The histone deacetylase inhibitor may be selected from the groupconsisting of Vorinostat, Romidepsin, Chidamide, Panobinostat,Belinostat, Valproic acid, Givinostat, and combinations thereof.

The anti-infective agent may be selected from the group consisting oforitavancin (Orbactiv), dalvavancin (Dalvance), tedizolid phosphate,(Sivextro), clindamycin, linezolid (Zyvox), mupirocin (Bactroban),trimethoprim, sulfamethoxazole, trimethoprim-sulfamethoxazole (Septra orBactrim), a tetracycline, vancomycin, daptomycin, fluoroquinolines, andcombinations thereof.

The bromodomain inhibitor may be selected from the group consisting ofOTX015/MK-8628, CPI-0610, BMS-986158, ZEN003694, GSK2820151, GSK525762,INCB054329, INCB057643, ODM-207, R06870810, BAY1238097, CC-90010,AZD5153, FT-1101, ABBV-744, RVX-000222, and combinations thereof.

Experimental Section Scheme 1

A synthesis of G-1 is described in Org. Biomol. Chem., 2010, 8,2252-2259, which is hereby incorporated by reference, and depicted inScheme 1. A catalytic amount of Sc(OTf)₃ (0.492 g, 1.0 mmol) inanhydrous acetonitrile (2.0 cm³) was added to the mixture of6-bromopiperonal (2.30 g, 10.0 mmol), p-aminoacetophenone (1.30 g, 10.0mmol) and cyclopentadiene (3.30 g, 50.0 mmol) in acetonitrile (25 cm³).The reaction mixture was stirred at ambient temperature (˜23° C.) for2.0 h. The volatiles were removed in vacuo. The residue was purified bypreparative silica gel column chromatography using ethyl acetate-hexanes(10:90) to provide G-1 (4.03 g, 98%, d.r.=94:6) as a white solid. Theminor diastereomer was substantially removed by recrystallization toyield a racemic mixture of SRR G-1 and RSS G-1.

Example 1: Isolation of the SRR G-1 and RSS G-1 Enantiomers

Starting with a highly purified sample of G-1,014446-bromobenzo[d][1,3]dioxol-5-yl)-(3aS*,4R*,9bR*)-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-one,(99.4% purity) purchased from Tocris Bioscience, the material wasdissolved in 90:10:0.1 (v/v/v) methyl tert-butyl ether/ethanol/diethylamine and subjected to preparative chromatography using a column packedwith Chrialpak 1A resin. Elution was conducted with 90:10:0.1 (v/v/v)methyl tert-butyl ether/ethanol/diethyl amine and the fractionscorresponding to each enantiomer were collected and concentered to asolid. The early eluting enantiomer was determined to be the SRR G-1enantiomer by single crystal x-ray structural analysis.

Example 2: SRR G-1 Polymorph Screen

Starting with SRR G-1 prepared according to Example 1, a polymorphscreening study was conducted analyzing the solids isolated from slurryof the solid, or from fast and slow evaporation and cooling of solutions(Table 1). Two crystal forms were identified, an anhydrous formdesignated Form A and mono dichloromethane solvate designated Form B. Onexposure to elevated temperature the Form B crystal form desolvates toform the Form C crystal form. Amorphous material was generated frompurified SRR G-1 by two different methods; quick evaporating a diethylether solution of SRR G-1 or rotary evaporating from a solution of adichloromethane solution of SRR G-1.

TABLE 1 Solvent Method¹ Observation² Result acetone fast evaporationwhite, blades and Form aciculars, B A + peak @6.9° 1. slowevaporation 1. crystals in tacky film Form A 2. scratched/mixed 2.nucleated, fines, B slow cool fine blades, aciculars, Form A B ACN fastevaporation white, aciculars, B Form A slow cool rosettes of blades, BForm A slurry, ambient, 14 d nucleated on stir bar, Form A aciculars, B,left wet added 88:12 H₂O/ACN clear, then precipitated Form A DCMrecrystallization clear, then precipitated, Form B blades, B fastevaporation blades, B Form B 1. rotary evap 1. foam — 2. scraped 2. freeflowing, NB diethyl fast evaporation glass, NB — ether evaporation w/N₂fine rosettes Form A fast evaporation rosettes of aciculars, B Form Aslurry, ambient, 14 days — Form A EtOH fast evaporation white, fineaciculars, B Form A slurry, ambient, 14 days analyzed as wet cake Form Aslow cool aciculars and blades, B Form A analyzed as wet cake cooling ofsolution — Form A EtOAc drop of solvent added to dissolved, fine —cooled melt aciculars and blades, B 1. Fast evaporation 1. glassy NBForm A 2. scratched 2. aciculars, B 1. slow evaporation 1. glass NB FormA 2. scratched 2. aciculars, B MeOH fast evaporation white, aciculars, BForm A slow cool aggregates blades, B Form A slurry, ambient, 14 days —Form A IPA fast evaporation white, blades, B Form A 1. added water,55:45 1. clear solution Form A IPA/H₂O 2. lamellae, B 2. refrigerated, 1day 3. blades/tablets, B 4. freezer, 1 day 1. slurry on 100° C. plate 1.clear (25 mg/ml) Form A 2. seeded w/7615-09-02 2. seeds remained 3. slowcool 3. thin blades, B THF 1. fast evaporation 1. glass, NB Form A 2.scratched 2. opaque toluene 1. fast evaporation 1. tacky film Form A 2.scratched 2. blades, B water slurry, 53° C., 6 days white Form AIPA/water filtrate fine aciculars, B Form A 55:45 partial evaporationIPA/H₂O filtrate limited aciculars, B Form A 89:11 cooling of solutionACN/H₂O filtrate thin aciculars, B Form A 97:03 cooling of solution¹Times and temperatures are approximate unless noted. ²B = birefringentand NB = non birefringent when material viewed using polarized lightmicroscopy.

Single Crystal Structure Determination of SRR G-1 (Form B)

Starting with SRR G-1 prepared according to Example 1, a suitable singlecrystal was grown from dichlomethane solution and analyzed bysingle-crystal X-ray diffractometry. The structure was determinedsuccessfully.

A single crystal was generated from a solution of dichloromethane afteran evaporative step. A colorless plate having approximate dimensions of0.19×0.14×0.03 mm³, was mounted on a polymer loop in random orientation.Preliminary examination and data collection were performed on a RigakuSuperNova diffractometer, equipped with a copper anode microfocus sealedX-ray tube (Cu Kα λ=1.54184 Å) and a Dectris Pilatus3 R 200K hybridpixel array detector. Cell constants and an orientation matrix for datacollection were obtained from least-squares refinement using the settingangles of 6979 reflections in the range 3.4920°<θ<77.1910°. The spacegroup was determined by the program CRYSALISPRO to be P212121(international tables no. 19). The data were collected to a maximumdiffraction angle (2θ) of 155.132° at room temperature.

Frames were integrated with CRYSALISPRO. A total of 10119 reflectionswere collected, of which 4368 were unique. Lorentz and polarizationcorrections were applied to the data. The linear absorption coefficientis 5.144 mm⁻¹ for Cu Kα radiation. An empirical absorption correctionusing CRYSALISPRO was applied. Transmission coefficients ranged from0.676 to 1.000. Intensities of equivalent reflections were averaged. Theagreement factor for the averaging was 3.4% based on intensity.

The structure was solved by direct methods using SHELXT. The remainingatoms were located in succeeding difference Fourier syntheses. Thestructure was refined using S_(HELXL)-2014. Hydrogen atoms on SRR G-1were refined independently. The dichloromethane hydrogen atoms wereincluded in the refinement but restrained to ride on the atom to whichthey are bonded. The structure was refined in full-matrix least-squaresby minimizing the function:

Σw(|F _(o)|² −|F _(c)|²)²

where the weight, w, is defined as 1/[σ²(Fo2)+(0.0464P)²+(0.1905P)],where P=(Fo2+2Fc2)/3. Scattering factors were taken from the“International Tables for Crystallography”. Of the 4368 reflections usedin the refinements, only the reflections with intensities larger thantwice their uncertainty [I>2σ(I)], 4071, were used in calculating thefit residual, R. The final cycle of refinement included 334 variableparameters, 0 restraints, and converged with respective unweighted andweighted agreement factors of:

$R = {{\sum{{{F_{o} - F_{c}}}/{\sum F_{o}}}} = {{{0.0}348R_{w}} = {\sqrt{\left( {\sum{{w\left( {F_{o}^{2} - F_{c}^{2}} \right)}^{2}/{\sum{w\left( F_{o}^{2} \right)}^{2}}}} \right)} = 0.0905}}}$

The standard deviation of an observation of unit weight (goodness offit) was 1.07. The highest peak in the final difference Fourier had anelectron density of 0.398 e/Å³. The minimum negative peak had a value of−0.438 e/Å³.

Calculated X-ray Powder Diffraction (XRPD) Pattern. A calculated XRPDpattern was generated for Cu radiation using MERCURY and the atomiccoordinates, space group, and unit cell parameters from the singlecrystal structure. Atomic Displacement Ellipsoid and Packing Diagrams.The atomic displacement ellipsoid diagram was prepared using MERCURY.Atoms are represented by 50% probability anisotropic thermal ellipsoids.Packing diagrams and additional figures were generated with MERCURY.Hydrogen bonding is represented as dashed lines. Assessment of chiralcenters was performed with PLATON. Absolute configuration is evaluatedusing the specification of molecular chirality rules.

The crystal system is orthorhombic and the space group is P212121. Thecell parameters and calculated volume are: a=6.43156(10) Å, b=13.0752(2)Å, c=25.2941(4) Å, α=90°, β=90°, γ=90°, V=2127.09(6) Å³. Standarduncertainty is written in crystallographic parenthesis notation, e.g.0.123(4) is equivalent to 0.123±0.004. The formula weight is 497.20 gmol⁻¹ with Z=4, resulting in a calculated density of 1.553 g cm⁻³.Further details of the crystal data and crystallographic data collectionparameters are summarized in Table 2. The quality of the structureobtained is high, as indicated by the fit residual, R, of 0.0348(3.48%). R-factors in the range 2%-6% are quoted to be the most reliablydetermined structures.

TABLE 2 Crystal Data and Data Collection Parameters SRR G-1 (Form B)Empirical formula C₂₂H₂₀BrCl₂NO₃ Formula weight (g mol⁻¹) 497.20Temperature (K) 300.14(10) Wavelength (Å) 1.54184 Crystal systemorthorhombic Space group P2₁2₁2₁ Unit cell parameters a = 6.43156(10) Åα = 90° b = 13.0752(2) Å β = 90° c = 25.2941(4) Å γ = 90° Unit cellvolume (Å³) 2127.09(6) Cell formula units, Z 4 Calculated density (gcm⁻³) 1.553 Absorption coefficient (mm⁻¹) 5.144 F(000) 1008 Crystal size(mm³) 0.19 × 0.14 × 0.03 Reflections used for cell measurement 6979 θrange for cell measurement 3.4920°-77.1910° Total reflections collected10119 Index ranges −8 ≤ h ≤ 7; −13 ≤ k ≤ 16; −31≤/≤29 θ range for datacollection θ_(min) = 3.495°, θ_(max) = 77.566° Completeness to θ_(max)97.3% Completeness to θ_(full) = 67.684° 100% Absorption correctionmulti-scan Transmission coefficient range 0.676-1.000 Refinement methodfull matrix least-squares on F² Independent reflections 4368 [R_(int) =0.0340, R_(σ) = 0.0401] Reflections [I > 2σ(I)] 4071Reflections/restraints/parameters 4368/0/334 Goodness-of-fit on F² S =1.07 Final residuals [I > 2σ(I)] R = 0.0348, R_(w) = 0.0905 Finalresiduals [all reflections] R = 0.0375, R_(w) = 0.0924 Largest diff.peak and hole (e Å⁻³) 0.398, −0.438 Max/mean shift/standard uncertainty0.001/0.000 Absolute structure determination Flack parameter: −0.018(13)Hooft parameter: −0.020(11) Friedel coverage: 95%

An atomic displacement ellipsoid drawing of SSR G-1 dichloromethanesolvate is shown in FIG. 1. The molecule observed in the asymmetric unitof the single crystal structure is consistent with the proposedmolecular structure of the SSR enantiomer. The asymmetric unit shown inFIG. 1 contains one SSR G-1 molecule and one dichloromethane molecule.

Packing diagrams viewed along the a, b, and c crystallographic axes areshown in FIGS. 2-4 respectively. Hydrogen bonding from the amine to thecarbonyl on adjacent molecules results in a one dimensional hydrogenbond network along the b axis, shown in FIG. 5.

The absolute structure can be determined through an analysis ofanomalous X-ray scattering by the crystal. Anomalous scattering isassessed through the intensity differences between Friedel pairs. Forthe reflection data measured up to θ_(max) the Friedel coverage was 95%.A refined parameter x, known as the Flack parameter, encodes therelative abundance of the two components in an inversion twin. Thestructure contains a fraction 1−x of the model being refined, and x ofits inverse. Provided that a low standard uncertainty is obtained, theFlack parameter should be close to 0 if the solved structure is correct,and close to 1 if the inverse model is correct. The measured Flackparameter for the structure of SSR G-1 dichloromethane solvate shown inFIG. 1 is −0.018 with a standard uncertainty of 0.013, which indicatesstrong inversion-distinguishing power.

Additional information regarding the absolute structure can be assessedby applying Bayesian statistics to Bijvoet differences. This analysisprovides a series of probabilities for different hypotheses of theabsolute structure. This analysis results in the Hooft y parameter,which is interpreted in the same fashion as the Flack x parameter. Inaddition, this analysis results in three probabilities that the absolutestructure is either correct, incorrect or a racemic twin. For thecurrent data set the (Flack equivalent) Hooft y parameter is −0.020(11),the probability that the structure is correct is 1.000, and theprobability that the structure is either incorrect or a racemic twin areboth less than 10⁻²⁰⁰.

The absolute configuration is labeled in FIG. 6. This is consistent withthe configuration of SRR G-1.

FIG. 7 shows a calculated XRPD pattern of SRR G-1 dichloromethanesolvate, generated from the single crystal structure. The calculatedXRPD pattern is identical to that assigned to bulk samples identified asdisplaying the Form B XRPD pattern in the polymorph screening studysummarized in Table 1.

Tables of positional parameters and their estimated standard deviations,anisotropic displacement factor coefficients, bond distances, bondangles, hydrogen bonds and angles, and torsion angles are shown.

TABLE 3 Positional Parameters and Their Estimated Standard DeviationsAtom x y z U(eq) Br(1) 8808.8(8)  6559.3(3)  3207.2(2)  53.07(15) Cl(2)3763(3) 5448.9(16)  5638.0(8)  101.5(6)  Cl(1) −612(3) 5050.5(18) 5795.3(10) 115.1(7)  N(1) 5577(5) 3525(3) 3323.1(13) 42.3(7) O(1)1123(6) 5915(2) 4486.3(13) 57.7(8) O(3) 7888(6) −878(3) 2427.3(15) 67.7(10) O(2) 2219(6) 7574(3) 4337.0(17)  70.3(11) C(14) 8462(6)2312(3) 3345.0(14) 35.2(7) C(6) 5733(6) 5262(3) 3683.6(14) 35.7(8) C(5)6356(7) 6265(3) 3603.9(15) 42.0(8) C(9) 8562(6) 3976(3) 3875.8(14)36.6(7) C(13) 9775(6) 3030(3) 3681.4(16) 38.5(8) C(8) 6968(6) 4360(3)3475.9(15) 36.5(8) C(15) 6446(6) 2602(3) 3178.5(14) 36.9(7) C(19)9214(6) 1360(3) 3200.4(15) 37.9(7) C(7) 3929(7) 5085(3) 3984.5(15)40.6(8) C(20) 8852(8) −340(3) 2729.4(15) 45.7(8) C(4) 5279(8) 7111(3) 3803(2)  49.6(10) C(18) 8068(6)  680(3) 2887.3(15) 40.3(8) C(17)6087(8)  995(3) 2720.5(16) 47.6(9) C(2) 2893(6) 5911(3) 4185.9(17)42.4(9) C(16) 5301(7) 1925(3) 2862.0(17) 45.9(9) C(12) 10391(8)  2539(4) 4200(2)  52.3(11) C(3) 3551(8) 6899(3) 4097.3(18)  49.6(10) C(11)9271(8) 2875(4) 4592.1(18)  55.4(12) C(21) 10883(8)  −717(4)  2942(2) 51.7(10) C(1)  702(8) 6967(4)  4595(2)  64.2(14) C(10) 7705(8) 3657(4)4421.2(17)  51.9(11) C(22)  1737(10) 4558(4)  5593(3)  80.0(18) H(22A)1615 4330 5229 96 H(22B) 2074 3968 5809 96 H(7)  3510(90)  4370(40) 4053(19)   56(14) H(8)  7690(60)  4600(30)  3131(15)  26(9) H(19)10640(70)  1160(30)  3313(17)   42(12) H(16)  3900(100)  2170(40) 2756(19)   59(14) H(17)  5370(90)  550(40)  2500(20)   60(15) H(4) 5640(80)  7770(40)  3731(18)   54(14) H(10A)  6310(90)  3300(40) 4390(20)   59(13) H(13) 10940(80)  3220(30)  3471(18)   48(12) H(11) 9270(70)  2600(40)  4938(19)   46(12) H(21A) 10780(90)  −790(40) 3310(20)   63(16) H(12) 11490(90)  2050(40)  4245(19)   59(14) H(21B) 11400(100) −1260(40)  2760(20)   70(16) H(10B)  7530(90)  4180(40) 4680(20)   57(14) H(9)  9630(70)  4540(30)  3937(16)   40(12) H(1) 4510(80)  3720(30)  3112(18)   47(13) H(1A)  −980(140)  7110(60) 4440(30)  130(30) H(1B)   980(110)  7000(50)  4990(30)  100(20) H(21C) 12080(110)  −240(50)  2910(30)   90(20) Hydrogen atoms were refinedisotropically except H22A&B, which were included in calculation ofstructure factors but not refined

TABLE 4 Anisotropic Displacement Factor Coefficients Atom U₁₁ U₂₂ U₃₃U₂₃ U₁₃ U₁₂ Br(1) 49.9(2)  50.7(2)  58.7(2)  10.2(2)  5.2(2) −10.2(2) Cl(2) 85.3(10) 111.4(13)  107.8(13)  19.4(10) −34.4(11)  −23.5(11) Cl(1) 93.0(14) 107.4(15)  144.8(18)  −22.8(14)  21.2(13)  1.5(11) N(1)35.1(16) 42.2(17) 49.7(18) −8.6(15) −9.2(13)  1.1(13) O(1) 44.1(15)61.5(18) 67.4(19) −18.9(15)  14.3(17) −3.6(16) O(3)  73(2) 54.4(19) 76(2) −28.4(18)  −17.7(19)   3.1(17) O(2)  59(2) 50.7(19) 102(3) −27.2(19)   10(2)  6.8(16) C(14) 33.6(18) 34.7(16) 37.2(17)  0.1(13)−0.9(14) −5.6(14) C(6) 35.4(19) 37.4(18) 34.2(16) −0.8(14) −3.4(14)−1.5(14) C(5) 39.8(19) 40.6(19) 45.6(18)  2.9(14) −3.7(18) −2.3(16) C(9)34.4(18) 36.2(17) 39.4(17) −2.7(14) −2.7(16) −3.3(15) C(13) 32.3(18)38.2(19)  45(2) −4.7(16) −4.3(16) −3.8(15) C(8) 36.5(19) 35.0(18)38.0(18) −0.1(15) −2.9(15) −0.7(14) C(15) 34.8(17) 38.7(17) 37.1(16) 0.0(14) −0.4(17) −1.7(14) C(19) 36.7(19) 38.9(18) 38.2(16) −1.3(15) 0.7(15) −1.4(13) C(7) 37.3(19) 39.1(19) 45.5(19) −2.9(15)  0.8(18)−2.3(17) C(20)  53(2) 41.3(19) 42.6(19) −6.0(15)  2(2)  −9(2) C(4) 52(2)  31(2)  65(3) −0.4(19)  −4(2) −3.5(17) C(18)  43(2)  41(2)37.2(18) −5.5(15) −0.6(16) −7.2(15) C(17)  44(2)  51(2)  48(2)−14.4(17)   −6(2)  −6(2) C(2) 35.5(19)  45(2)  47(2) −9.1(17)  0.6(17)−0.1(16) C(16)  40(2)  51(2)  47(2) −13.0(18)  −10.0(18)   0.9(17) C(12) 50(2)  46(2)  61(3)  −7(2) −26(2)   2.7(19) C(3)  47(2)  44(2)  58(2)−13.6(18)   −5(2)  5.8(18) C(11)  68(3)  55(3)  43(2)  1.7(19) −17(2)  3(2) C(21)  54(3)  45(2)  57(3) −10.2(19)   0(2)  4.0(19) C(1)  50(3) 71(3)  72(3) −28(3)   5(2)  11(2) C(10)  60(3)  60(3) 35.4(19)  1.5(19)−4.8(19)  7(2) C(22)  86(4)  52(3) 103(4)  −11(3)  −27(4)   10(3) Theform of the anisotropic temperature factor is: exp[−2□ h²a*²U(1,1) +k²b*²U(2,2) + l²c*²U(3,3) + 2hka*b*U(1,2) + 2hla*c*U(1,3) +2klb*c*U(2,3)] where a*, b*, and c* are reciprocal lattice constants.

TABLE 5 Bond Distances in Ångströms Atom Atom Length/Å Atom AtomLength/Å Br(1) C(5) 1.909(4) C(5) C(4) 1.399(6) Cl(2) C(22) 1.751(6)C(9) C(13) 1.543(5) Cl(1) C(22) 1.720(7) C(9) C(8) 1.525(5) N(1) C(8)1.463(5) C(9) C(10) 1.543(6) N(1) C(15) 1.380(5) C(13) C(12) 1.514(6)O(1) C(2) 1.369(5) C(15) C(16) 1.402(5) O(1) C(1) 1.430(6) C(19) C(18)1.400(5) O(3) C(20) 1.209(5) C(7) C(2) 1.367(6) O(2) C(3) 1.371(5) C(20)C(18) 1.481(6) O(2) C(1) 1.417(7) C(20) C(21) 1.495(7) C(14) C(13)1.523(5) C(4) C(3) 1.366(7) C(14) C(15) 1.415(5) C(18) C(17) 1.404(7)C(14) C(19) 1.385(5) C(17) C(16) 1.365(6) C(6) C(5) 1.385(5) C(2) C(3)1.378(6) C(6) C(8) 1.516(5) C(12) C(11) 1.302(7) C(6) C(7) 1.407(6)C(11) C(10) 1.499(7) Numbers in parentheses are estimated standarddeviations in the least significant digits.

TABLE 6 Bond Angles in Degrees Atom Atom Atom Angle/° Atom Atom AtomAngle/° C(15) N(1) C(8) 118.4(3) C(16) C(15) C(14) 118.8(3) C(2) O(1)C(1) 105.6(4) C(14) C(19) C(18) 122.4(4) C(3) O(2) C(1) 105.9(4) C(2)C(7) C(6) 118.3(4) C(15) C(14) C(13) 120.6(3) O(3) C(20) C(18) 121.3(4)C(19) C(14) C(13) 120.6(3) O(3) C(20) C(21) 118.9(4) C(19) C(14) C(15)118.8(3) C(18) C(20) C(21) 119.8(4) C(5) C(6) C(8) 122.3(4) C(3) C(4)C(5) 116.0(4) C(5) C(6) C(7) 118.2(4) C(19) C(18) C(20) 123.0(4) C(7)C(6) C(8) 119.4(3) C(19) C(18) C(17) 117.5(4) C(6) C(5) Br(1) 120.4(3)C(17) C(18) C(20) 119.5(4) C(6) C(5) C(4) 123.6(4) C(16) C(17) C(18)121.3(4) C(4) C(5) Br(1) 116.0(3) O(1) C(2) C(3) 110.0(4) C(8) C(9)C(13) 113.1(3) C(7) C(2) O(1) 128.0(4) C(8) C(9) C(10) 116.2(3) C(7)C(2) C(3) 122.0(4) C(10) C(9) C(13) 104.4(3) C(17) C(16) C(15) 121.1(4)C(14) C(13) C(9) 113.1(3) C(11) C(12) C(13) 111.8(4) C(12) C(13) C(14)111.6(3) O(2) C(3) C(2) 109.8(4) C(12) C(13) C(9) 101.3(3) C(4) C(3)O(2) 128.2(4) N(1) C(8) C(6) 110.6(3) C(4) C(3) C(2) 121.9(4) N(1) C(8)C(9) 109.9(3) C(12) C(11) C(10) 112.5(4) C(6) C(8) C(9) 112.2(3) O(2)C(1) O(1) 108.6(4) N(1) C(15) C(14) 121.8(3) C(11) C(10) C(9) 101.7(4)N(1) C(15) C(16) 119.4(3) Cl(1) C(22) Cl(2) 112.7(3) Numbers inparentheses are estimated standard deviations in the least significantdigits.

TABLE 7 Hydrogen Bond Distances in Ångströms and Angles in Degrees DonorH Acceptor D-H H . . . A D . . . A D-H . . . A N(1) H(1) O(3) 0.91 (5)2.13(5) 3.030(5) 176(4) Numbers in parentheses are estimated standarddeviations in the least significant digits.

TABLE 8 Torsion Angles in Degrees A B C D Angle/° A B C D Angle/° Br(1)C(5) C(4) C(3) −178.5(3) C(8) C(9) C(10) C(11) 152.3(4) N(1) C(15) C(16)C(17) −178.5(4) C(15) N(1) C(8) C(6) −173.2(3) O(1) C(2) C(3) O(2)−0.2(5) C(15) N(1) C(8) C(9) −48.7(5) O(1) C(2) C(3) C(4) −179.4(4)C(15) C(14) C(13) C(9) 8.6(5) O(3) C(20) C(18) C(19) −174.5(4) C(15)C(14) C(13) C(12) 122.0(4) O(3) C(20) C(18) C(17) 5.6(6) C(15) C(14)C(19) C(18) 1.2(6) C(14) C(13) C(12) C(11) −102.7(5) C(19) C(14) C(13)C(9) −170.9(3) C(14) C(15) C(16) C(17) 0.9(6) C(19) C(14) C(13) C(12)−57.5(5) C(14) C(19) C(18) C(20) 180.0(4) C(19) C(14) C(15) N(1)177.9(3) C(14) C(19) C(18) C(17) −0.2(6) C(19) C(14) C(15) C(16) −1.6(5)C(6) C(5) C(4) C(3) 1.2(7) C(19) C(18) C(17) C(16) −0.5(6) C(6) C(7)C(2) O(1) 180.0(4) C(7) C(6) C(5) Br(1) 179.2(3) C(6) C(7) C(2) C(3)0.8(6) C(7) C(6) C(5) C(4) −0.5(6) C(5) C(6) C(8) N(1) −146.0(4) C(7)C(6) C(8) N(1) 36.3(5) C(5) C(6) C(8) C(9) 90.8(4) C(7) C(6) C(8) C(9)−86.8(4) C(5) C(6) C(7) C(2) −0.5(6) C(7) C(2) C(3) O(2) 179.1(4) C(5)C(4) C(3) O(2) −179.9(5) C(7) C(2) C(3) C(4) 0.0(7) C(5) C(4) C(3) C(2)−0.9(7) C(20) C(18) C(17) C(16) 179.3(4) C(9) C(13) C(12) C(11) 17.9(5)C(18) C(17) C(16) C(15) 0.1(7) C(13) C(14) C(15) N(1) −1.6(5) C(2) O(1)C(1) O(2) 2.1(6) C(13) C(14) C(15) C(16) 179.0(4) C(12) C(11) C(10) C(9)−17.1(6) C(13) C(14) C(19) C(18) −179.3(4) C(3) O(2) C(1) O(1) −2.2(6)C(13) C(9) C(8) N(1) 54.2(4) C(21) C(20) C(18) C(19) 4.9(6) C(13) C(9)C(8) C(6) 177.7(3) C(21) C(20) C(18) C(17) −174.9(4) C(13) C(9) C(10)C(11) 27.0(4) C(1) O(1) C(2) C(7) 179.5(5) C(13) C(12) C(11) C(10)−0.5(6) C(1) O(1) C(2) C(3) −1.2(5) C(8) N(1) C(15) C(14) 22.9(5) C(1)O(2) C(3) C(4) −179.4(5) C(8) N(1) C(15) C(16) −157.7(4) C(1) O(2) C(3)C(2) 1.5(6) C(8) C(6) C(5) Br(1) 1.5(5) C(10) C(9) C(13) C(14) 92.4(4)C(8) C(6) C(5) C(4) −178.2(4) C(10) C(9) C(13) C(12) −27.1(4) C(8) C(6)C(7) C(2) 177.3(4) C(10) C(9) C(8) N(1) −66.7(4) C(8) C(9) C(13) C(14)−34.9(4) C(10) C(9) C(8) C(6) 56.9(5) C(8) C(9) C(13) C(12) −154.4(3)Numbers in parentheses are estimated standard deviations in the leastsignificant digits.

Starting with SRR G-1 prepared according to Example 1, a suitable singlecrystal of the Form A crystal form was grown from isopropanol solutionand analyzed by single-crystal X-ray diffractometry. The structure wasdetermined successfully. A single crystal x-ray analysis was conductedon a colorless plate having approximate dimensions of 0.203×0.137×0.033mm³, was mounted on a polymer loop in random orientation. Preliminaryexamination and data collection were performed on a Rigaku SuperNovadiffractometer, equipped with a copper anode microfocus sealed X-raytube (Cu Kα λ=1.54184 Å) and a Dectris Pilatus3 R 200K hybrid pixelarray detector. Cell constants and an orientation matrix for datacollection were obtained from least-squares refinement using the settingangles of 9009 reflections in the range 4.7640°<θ<77.3860°. The spacegroup was determined by the program CRYSALISPRO to be P2₁2₁2₁. The datawere collected to a maximum diffraction angle (2θ) of 155.264° at roomtemperature.

Frames were integrated with CRYSALISPRO. A total of 17299 reflectionswere collected, of which 7561 were unique. Lorentz and polarizationcorrections were applied to the data. The linear absorption coefficientis 3.206 mm⁻¹ for Cu Kα radiation. An empirical absorption correctionusing CRYSALISPRO was applied. Transmission coefficients ranged from0.733 to 1.000. Intensities of equivalent reflections were averaged. Theagreement factor for the averaging was 2.74% based on intensity.

The structure was solved by direct methods using SHELXT. The remainingatoms were located in succeeding difference Fourier syntheses. Thestructure was refined using SHELXL-2014. Hydrogen atoms were refinedindependently. The structure was refined in full-matrix least-squares byminimizing the function:

Σw(|F _(o)|² −|F _(c)|²)²

where the weight, w, is defined as 1/[σ²(F_(o) ²)+(0.0401P)²+(0.3205P)],where P=(F_(o) ²+2F_(c) ²)/3. Scattering factors were taken from the“International Tables for Crystallography”. Of the 7561 reflections usedin the refinements, only the reflections with intensities larger thantwice their uncertainty [I>2σ(I)], 6752, were used in calculating thefit residual, R. The final cycle of refinement included 613 variableparameters, 0 restraints, and converged with respective unweighted andweighted agreement factors of:

R = ∑F_(o) − F_(c)/∑F_(o) = 0.0325$R_{w} = {\sqrt{\left( {\sum{{w\left( {F_{o}^{2} - F_{c}^{2}} \right)}^{2}/{\sum{w\left( F_{o}^{2} \right)}^{2}}}} \right)} = {0.0813}}$

The standard deviation of an observation of unit weight (goodness offit) was 1.04. The highest peak in the final difference Fourier had anelectron density of 0.319 e/Å³. The minimum negative peak had a value of−0.454 e/Å³.

The crystal system is orthorhombic and the space group is P2₁2₁2₁. Thecell parameters and calculated volume are: a=6.50106(9) Å, b=17.3547(2)Å, c=32.6957(4) Å, α=90°, β=90°, γ=90°, V=3688.85(9) Å³. The molecularweight is 412.27 g mol⁻¹ with Z=8, resulting in a calculated density of1.485 g cm⁻³. Further details of the crystal data and crystallographicdata collection parameters are summarized in Table 9. An atomicdisplacement ellipsoid drawing of Form A is shown in FIG. 8. Theasymmetric unit shown contains two enantiopure SRR G-1 molecules. Fromthe structure, the absolute configuration was determined conclusively.SRR G-1 contains three chiral centers located at C114 (C214), C113(C213), and C19 (C29) which bond in the R, S, and R configuration,respectively. A calculated XRPD pattern of Form A, generated from thesingle crystal structure, is provided in FIG. 9 and compared to theexperimental pattern.

TABLE 9 Crystal Data and Data Collection Parameters for SRR G-1 Form AEmpirical formula C₂₁H₁₈BrNO₃ Formula weight (g mol⁻¹) 412.27Temperature (K) 299.84(10) Wavelength (Å) 1.54184 Crystal systemorthorhombic Space group P2₁2₁2₁ Unit cell parameters a = 6.50106(9) Å α= 90° b = 17.3547(2) Å β = 90° c = 32.6957(4) Å γ = 90° Unit cell volume(Å³) 3688.85(9) Cell formula units, Z 8 Calculated density (g cm⁻³)1.485 Absorption coefficient (mm⁻¹) 3.206 F(000) 1680 Crystal size (mm³)0.203 × 0.137 × 0.033 Reflections used for cell measurement 9009 θ rangefor cell measurement 4.7640°-77.3860° Total reflections collected 17299Index ranges −8 ≤ h ≤ 7; −21 ≤ k ≤ 10; −40≤/≤38 θ range for datacollection θ_(min) = 3.714°, θ_(max) = 77.632° Completeness to θ_(max)98.3% Completeness to θ_(full) = 67.684° 99.9% Absorption correctionmulti-scan Transmission coefficient range 0.733-1.000 Refinement methodfull matrix least-squares on F² Independent reflections 7561 [R_(int) =0.0274, R_(σ) = 0.0337] Reflections [I > 2σ(I)] 6752Reflections/restraints/parameters 7561/0/613 Goodness-of-fit on F² S =1.04 Final residuals [I > 2σ(I)] R = 0.0325, R_(w) = 0.0813 Finalresiduals [all reflections] R = 0.0373, R_(w) = 0.0843 Largest diff.peak and hole (e Å⁻³) 0.319, −0.454 Max/mean shift/standard uncertainty0.002/0.000 Absolute structure determination Flack parameter: −0.015(9)

Example 3: Form and Polymorph Data

SRR G-1 forms two distinctive polymorphs, Forms A and C; a solvate, FormB; as well as amorphous material. The XRPD patterns for the crystallineforms are compared in FIG. 10. Form B is a mono DCM solvate thatdesolvates to Form C upon exposure to elevated temperatures between 100and 120° C. Form C, the desolvate, exhibits a melt onset near 129° C.Form A is the thermodynamically stable form at all temperatures(monotropically related to Form C) and exhibits a melt onset near 178°C. Amorphous material is not physically stable and crystallizes to FormA upon exposure to either elevated temperature or humidity. The formsare discussed in more detail in subsequent sections below.

Crystalline Form A

Form A is anhydrous with a melt onset near 178° C. Form A isthermodynamically the most stable form, relative, monotropically, toForm C. Form A was routinely obtained from multiple crystallizationtechniques utilizing various organic solvents and organic/water solventsystems other than dichloromethane.

The observed XRPD peaks for Crystalline Form A are listed in Table 10

TABLE 10 Diffraction angle 2θ (°) d-spacing (Å) Intensity (%)  5.39 ±0.20 16.387 ± 0.608  19  5.75 ± 0.20 15.364 ± 0.534  100  9.56 ± 0.209.245 ± 0.193 43 10.17 ± 0.20 8.689 ± 0.170 25 10.53 ± 0.20 8.397 ±0.159 60 10.81 ± 0.20 8.181 ± 0.151 30 11.52 ± 0.20 7.675 ± 0.133 1411.95 ± 0.20 7.400 ± 0.123 4 13.02 ± 0.20 6.795 ± 0.104 31 13.88 ± 0.206.377 ± 0.091 7 14.66 ± 0.20 6.036 ± 0.082 34 14.79 ± 0.20 5.985 ± 0.08036 15.52 ± 0.20 5.705 ± 0.073 20 15.87 ± 0.20 5.578 ± 0.070 5 16.23 ±0.20 5.457 ± 0.067 43 16.67 ± 0.20 5.315 ± 0.063 10 17.03 ± 0.20 5.204 ±0.061 70 17.23 ± 0.20 5.142 ± 0.059 9 17.88 ± 0.20 4.958 ± 0.055 1118.16 ± 0.20 4.882 ± 0.053 5 18.89 ± 0.20 4.695 ± 0.049 28 19.22 ± 0.204.614 ± 0.048 23 19.91 ± 0.20 4.456 ± 0.044 12 20.22 ± 0.20 4.389 ±0.043 15 20.54 ± 0.20 4.321 ± 0.042 80 20.71 ± 0.20 4.285 ± 0.041 7621.25 ± 0.20 4.178 ± 0.039 86 21.86 ± 0.20 4.062 ± 0.037 88 22.10 ± 0.204.019 ± 0.036 9 22.39 ± 0.20 3.968 ± 0.035 11 23.44 ± 0.20 3.793 ± 0.03214 23.62 ± 0.20 3.763 ± 0.031 24 23.99 ± 0.20 3.706 ± 0.030 17 24.67 ±0.20 3.606 ± 0.029 60 25.25 ± 0.20 3.524 ± 0.027 23 25.61 ± 0.20 3.475 ±0.027 28 25.99 ± 0.20 3.425 ± 0.026 9 26.27 ± 0.20 3.390 ± 0.025 3026.94 ± 0.20 3.307 ± 0.024 16 27.24 ± 0.20 3.271 ± 0.024 6 28.06 ± 0.203.177 ± 0.022 44 29.13 ± 0.20 3.063 ± 0.021 13 29.33 ± 0.20 3.042 ±0.020 20 29.66 ± 0.20 3.009 ± 0.020 21 30.04 ± 0.20 2.972 ± 0.019 13

Thermograms of Form A are shown in FIG. 11. Thermogravimetric Analysis(TGA) data shows no weight loss up to 266° C., consistent with ananhydrous form. The DSC exhibits a single endotherm with an onset near176° C. (68 J/g). The event was visually confirmed on a hot plate as amelt. Discoloration, likely due to decomposition, was noted uponmelting.

The Dynamic Vapor Sorption isotherm for Form A indicates the formexhibits low hygroscopicity (FIG. 12). The weight change through thesorption/desorption cycle was less than 0.3% weight. Hysteresis was notobserved. The material recovered from the Dynamic Vapor Sorptionexperiment was Form A by XRPD.

Crystalline Form B

Form B is monodichloromethane solvate generated routinely as a mixturewith Form C (desolvated form) from DCM. Form B will desolvate fully toForm C when exposed to temperatures between 100 and 120° C.

The single crystal structure for Form B is known. The crystal system isorthorhombic and the space group is P2₁2₁2₁. The cell parameters andcalculated volume are: a=6.43156(10) Å, b=13.0752(2) Å, c=25.2941(4) Å,α=90°, β=90°, γ=90°, V=2127.09(6) Å³. The formula weight is 497.20 gmol⁻¹ with Z=4, resulting in a calculated density of 1.553 g cm⁻³. Theasymmetric unit contains one enantiopure SRR G-1 molecule and onedichloromethane molecule. The structure contains three chiral centerslocated at C8, C9, and C13 (refer to FIG. 13) which bond in the R, S,and R configuration, respectively. A calculated XRPD pattern of Form B,generated from the single crystal structure, is provided in FIG. 14 andcompared to the experimental pattern.

The observed XRPD peaks for Crystalline Form B are listed in Table 11

TABLE 11 Diffraction angle 2θ (°) d-spacing (Å) Intensity (%)  6.97 ±0.20 12.677 ± 0.363  7  7.60 ± 0.20 11.627 ± 0.306  25  9.71 ± 0.209.097 ± 0.187 25 13.53 ± 0.20 6.540 ± 0.096 6 13.98 ± 0.20 6.331 ± 0.09075 14.19 ± 0.20 6.236 ± 0.087 33 15.44 ± 0.20 5.735 ± 0.074 64 15.73 ±0.20 5.628 ± 0.071 11 16.87 ± 0.20 5.251 ± 0.062 9 17.14 ± 0.20 5.168 ±0.060 3 17.33 ± 0.20 5.114 ± 0.059 3 18.61 ± 0.20 4.764 ± 0.051 24 19.36± 0.20 4.582 ± 0.047 3 19.67 ± 0.20 4.510 ± 0.045 44 20.64 ± 0.20 4.299± 0.041 10 20.82 ± 0.20 4.262 ± 0.040 28 21.06 ± 0.20 4.216 ± 0.040 321.55 ± 0.20 4.119 ± 0.038 100 22.05 ± 0.20 4.027 ± 0.036 57 23.36 ±0.20 3.806 ± 0.032 6 23.96 ± 0.20 3.712 ± 0.031 12 24.65 ± 0.20 3.608 ±0.029 30 24.91 ± 0.20 3.572 ± 0.028 9 25.11 ± 0.20 3.544 ± 0.028 1225.66 ± 0.20 3.469 ± 0.027 15 26.18 ± 0.20 3.401 ± 0.026 29 26.86 ± 0.203.317 ± 0.024 7 27.50 ± 0.20 3.241 ± 0.023 24 27.71 ± 0.20 3.217 ± 0.0237 27.94 ± 0.20 3.191 ± 0.022 15 28.18 ± 0.20 3.164 ± 0.022 32 28.54 ±0.20 3.125 ± 0.021 13 28.75 ± 0.20 3.103 ± 0.021 15 29.03 ± 0.20 3.073 ±0.021 4 29.44 ± 0.20 3.031 ± 0.020 20 29.70 ± 0.20 3.005 ± 0.020 7

The thermograms for Form B are shown in FIG. 15. The ThermogravametricAnalysis (TGA) curve exhibits a weight loss of approximately 15.3% up to177° C., consistent with the volatilization of 0.9 mol/mol DCM. The lossoccurs concurrently with a desolvation endotherm (max 104° C.) andrecrystallization exotherm (max 140° C.) in the DSC thermogram. Therecrystallized material exhibits a final melt endotherm with an onsetnear 176° C. consistent with the melt of Form A. The DSC thermogram forthe mixture of Forms B and C is provided in FIG. 16. The mixtureexhibits a desolvation endotherm (max 101° C.) followed by the meltendotherm (onset near 128° C.) of the desolvated form, Form C.

The physical stability of Form B was investigated. Complete desolvationto Form C occurred upon exposure to 120° C. Almost complete desolvationwas evident upon exposure to 90 to 100° C. for 25 minutes. Vacuum at 70°C. (or below) was not sufficient for desolvation to occur.

Crystalline Form C

Form C is a desolvate with a melt onset near 129° C. generated throughthe desolvation of Form B (mono DCM solvate).

The XRPD pattern of Form C was successfully indexed, suggesting it iscomposed of a single crystalline phase (FIG. 17). Assuming the chemicalcomposition is correct, it has an orthorhombic unit cell containing fourmolecules of SRR G-1. Consequently, the formula unit volume of 462.88 Å³calculated from the indexing results would be consistent with ananhydrous form with a calculated density of 1.479 g cm⁻³.

The observed XRPD peaks for Crystalline Form C are listed in Table 12

TABLE 12 Diffraction angle 2θ (°) d-spacing (Å) Intensity (%)  7.69 ±0.20 11.483 ± 0.298  65  8.62 ± 0.20 10.250 ± 0.237  42 10.73 ± 0.208.235 ± 0.153 78 12.77 ± 0.20 6.925 ± 0.108 97 13.49 ± 0.20 6.560 ±0.097 100 14.22 ± 0.20 6.222 ± 0.087 30 14.99 ± 0.20 5.906 ± 0.078 3315.60 ± 0.20 5.674 ± 0.072 22 16.09 ± 0.20 5.506 ± 0.068 74 17.32 ± 0.205.117 ± 0.059 34 18.24 ± 0.20 4.860 ± 0.053 26 19.17 ± 0.20 4.626 ±0.048 18 19.71 ± 0.20 4.500 ± 0.045 39 19.86 ± 0.20 4.466 ± 0.045 6820.60 ± 0.20 4.308 ± 0.041 82 21.10 ± 0.20 4.206 ± 0.039 35 22.05 ± 0.204.028 ± 0.036 71 22.78 ± 0.20 3.900 ± 0.034 33 22.98 ± 0.20 3.868 ±0.033 62 23.59 ± 0.20 3.768 ± 0.031 19 24.00 ± 0.20 3.706 ± 0.030 3625.16 ± 0.20 3.536 ± 0.028 27 25.57 ± 0.20 3.481 ± 0.027 20 26.09 ± 0.203.413 ± 0.026 44 26.46 ± 0.20 3.366 ± 0.025 16 27.01 ± 0.20 3.299 ±0.024 22 27.30 ± 0.20 3.264 ± 0.023 14 28.41 ± 0.20 3.139 ± 0.022 1328.76 ± 0.20 3.102 ± 0.021 15 28.90 ± 0.20 3.087 ± 0.021 16 29.18 ± 0.203.058 ± 0.021 24 29.43 ± 0.20 3.032 ± 0.020 23 30.23 ± 0.20 2.954 ±0.019 39

The differential scanning calorimetry (DSC) thermogram for Form C isshown in FIG. 18. The DSC exhibits a single endotherm with an onset near129° C. (23 J/g). The event was visually confirmed on a hot plate as amelt.

Amorphous

The physical stability of amorphous material generated from purified SRRG-1 was investigated. Amorphous material crystallized to Form A uponexposure to either elevated temperature (within 4 days at 60° C.) orhumidity (within 12 days at 75% RH). This indicates that amorphousmaterial is not stable at the conditions evaluated.

Relative Thermodynamic Stability of the Crystalline Forms

Phase transitions of solids can be thermodynamically reversible orirreversible. Crystalline forms which transform reversibly at a specifictransition temperature are called enantiotropic polymorphs. If thecrystalline forms are not interconvertible under these conditions, thesystem is monotropic (one thermodynamically stable form). Several ruleshelp predict the relative thermodynamic stability of polymorphs andwhether the relationship between the polymorphs is enantiotropic ormonotropic. The density and heat of fusion rules, justified on astatistical mechanical basis, are used here for guidance of monotropy orenantiotropy.

The density rule, which is based on Kita{hacek over (i)}gorodski{hacekover (i)}'s principle of closest packing for molecular crystals, statesthat, for a non-hydrogen-bonded system at absolute zero, the most stablepolymorph will have the highest density, because of strongerintermolecular van der Waals interactions. Thus, according to this rule,the crystal structure with most efficient packing will also have thelowest free energy. This assumes that hydrogen bonding (long rangeeffect) is not a major parameter in crystal packing. The densitiesdetermined from the single crystal structure of Form A and indexingresults of Form C suggest that, at absolute zero, Form A is more stablethan Form C (1.485 and 1.479 g cm⁻³, respectively).

The melt onsets and heats of fusion, obtained from calorimetry data, areuseful to estimate the relative physical stabilities of the forms at alltemperatures (FIGS. 11 and 18). From the heat of fusion rule, two formsare enantiotropic if the higher melting form has the lower heat offusion, otherwise they are monotropic. The density and heat of fusionrules for this system appear consistent with a monotropic relationship.

Interconversion experiments were performed to experimentally test thethermodynamic relationship between Forms A and C. Interconversion orcompetitive slurry experiments are a solution-mediated process thatprovides a pathway for the less soluble (more stable) crystal to grow atthe expense of the more soluble crystal form. Outside the formation of asolvate or degradation, the resulting more stable polymorph from aninterconversion experiment is independent of the solvent used becausethe more thermodynamically stable polymorph has a lower energy andtherefore lower solubility. The choice of solvent affects the kineticsof polymorph conversion and not the thermodynamic relationship betweenpolymorphic forms. Saturated solutions were generated and then added tomixtures composed of approximately equivalent quantities of the twopolymorphs. The samples were slurried for nine days and the solidsharvested and analyzed by XRPD. The results of the interconversionstudies confirm Form A is thermodynamically more stable at roomtemperature relative to Form C. The experimentally determined relativestability at room temperature, the suggested relative stability atabsolute zero based on the density rule, and monotropism as determinedby the heat of fusion rule all imply that Form A is more stable thanForm C at any temperature.

Solubility of SRR G-1 Form A

TABLE 13 Approximate Solubility of Purified SRR G-1 Form A SolventSolubility (mg/mL) acetone >122 ACN 24 DCM >89 DMSO >203 diethyl ether 8EtOH 7 EtOAc >65 IPA 4 MeOH 6 sesame oil 2 THF >67 toluene 40 water <2

SRR G-1 (Form A) Solubility Measurement in pH Buffers

Solubility measurement was performed for SRR G-1 (crystal Form A) in pHbuffers (2.0˜8.0) at 37° C. for 24 hrs. The results were summarized inTable 14. The XRPD patterns are shown in FIG. 19 and FIG. 20. No formchange was observed for the compound in all the pH buffers after 24 hrs.The solubility at pH 2.0˜8.0 for SRR G-1 was less than 0.72 μg/mL.

TABLE 14 Summary of solubility measurement of SRR G-1 (Form A) in pHbuffers Form Media Temp. Solubility Final pH Conversion pH 2.0 37° C.<0.72 μg/mL 2.0 No pH 3.0 <0.72 μg/mL 3.0 No pH 4.0 <0.72 μg/mL 3.8 NopH 5.0 <0.72 μg/mL 5.1 No pH 6.0 <0.72 μg/mL 6.1 No pH 7.0 <0.72 μg/mL7.1 No pH 8.0 <0.72 μg/mL 8.0 No

SRR G-1 (Form A) Solubility Measurement in BioRelovent Media

Kinetic solubility measurement was performed for SRR G-1 (crystal FormA) in three bio-relevant media (SGF (pH 1.8), FaSSIF (pH 6.5) and FeSSIF(pH 5.0)) at 37° C. for 1, 2, 4 and 24 hrs. The results were summarizedin Table 15 and FIG. 21. The XRPD patterns of the wetcake are shown inFIGS. 22-24. No form change was observed for the sample after 1, 2, 4and 24 hrs in the three bio-relevant media. The highest solubility ofSRR G-1 was observed in FeSSIF (˜0.037 mg/mL).

TABLE 15 Summary of solubility measurement in bio-relevant media SGFFaSSIF FeSSIF Time Point Form Solubility Solubility Solubility (h)Change (mg/mL) pH FC (mg/mL) pH FC (mg/mL) pH 1 No 0.0059 1.9 No 0.00716.4 No 0.036 5.0 2 No 0.0062 1.9 No 0.0068 6.4 No 0.037 5.0 4 No 0.00661.9 No 0.0071 6.4 No 0.038 5.0 24 No 0.0057 1.7 No 0.0073 6.5 No 0.0375.0

The pKa, Log D_(7.4) and Log P of compound SRR G-1 were predicted byMarvinSketch 5.6.0.2, the results showed the pKa of SRR G-1 is 1.90(base, pH range of 0-14), Log D_(7.4) is 5.32 and Log P is 5.32. A pKatitration test showed that no pKa value was observed in the range of3˜11, which was consistent with the prediction result. Log D_(7.4) wasmeasured with the solvent systems of pH 7.4 phosphate buffer andn-octanol by shake-flask method. Detailed results of Log D_(7.4) and LogP were displayed in Table 16. Since the solubility of SRR G-1 freebasein aqueous phase was lower than <0.82 μg/mL, the Log D_(7.4) wasdetermined to be >3.22 and Log P was >3.22 considering the small pKavalue.

TABLE 16 LogD_(7.4) and LogP of compound SRR G-1 Concentration (mg/mL)Sample n- pH 7.4 Average of Simulated Simulated Name # octanol bufferLogD_(7.4) LogD_(7.4) LogD_(7.4) LogP* LogP SRR G-1 1 1.37 <0.82μg/mL >3.22 >3.22 5.32 >3.22 5.32 2 1.34 <0.82 μg/mL >3.21 3 1.37 <0.82μg/mL >3.22 *LogP = LogD_((pH)) + Log [1 + 10^((pKa −pH))]

Instrumental Techniques

Differential Scanning calorimetry (DSC)

DSC was performed using a Mettler-Toledo DSC3+ or DSC822e differentialscanning calorimeter. A tau lag adjustment is performed with indium,tin, and zinc. The temperature and enthalpy are adjusted with octane,phenyl salicylate, indium, tin and zinc. The adjustment is then verifiedwith octane, phenyl salicylate, indium, tin, and zinc. The sample wasplaced into a hermetically sealed aluminum DSC pan, and the weight wasaccurately recorded. The pan was then inserted into the DSC cell. Aweighed aluminum pan configured as the sample pan was placed on thereference side of the cell. The pan lid was pierced prior to sampleanalysis. Samples were analyzed from −30° C. to 250° C. @ 10°/min. Thecyclic DSC method heated from −30° C. to 100° C., returned to −30° C.,then heated to 250° C. at 10°/min.

Dynamic Vapor Sorption/Desorption (DVS)

Moisture sorption/desorption data were collected on a VTI SGA-100 VaporSorption Analyzer. NaCl and PVP were used as calibration standards.Samples were not dried prior to analysis. Sorption and desorption datawere collected over a range from 5% to 95% RH at 10% RH increments undera nitrogen purge. The equilibrium criterion used for analysis was lessthan 0.0100% weight change in 5 minutes with a maximum equilibrationtime of 3 hours. Data were not corrected for the initial moisturecontent of the samples.

Thermogravimetric Analysis (TGA)

Thermogravimetric analysis was performed using a Mettler-Toledo TGA/DSC3analyzer. Temperature and enthalpy adjustments were performed usingindium, tin, and zinc, and then verified with indium. The balance wasverified with calcium oxalate. The sample was placed in an open aluminumpan. The pan was hermetically sealed, the lid pierced, then insertedinto the TG furnace. A weighed aluminum pan configured as the sample panwas placed on the reference platform. The furnace was heated undernitrogen. Each sample was heated from ambient temperature to 350° C. at10° C./min. Although thermograms are plotted by reference temperature(x-axis), results are reported according to sample temperatures.

X-Ray Powder Diffraction (XRPD)

XRPD pattern was collected with a PANalytical X'Pert PRO MPD orPANalytical Empyrean diffractometer using an incident beam of Curadiation produced using a long, fine-focus source. An ellipticallygraded multilayer mirror was used to focus Cu Kα X-rays through thespecimen and onto the detector. Prior to the analysis, a siliconspecimen (NIST SRM 640e) was analyzed to verify the observed position ofthe Si 111 peak is consistent with the NIST-certified position. Aspecimen of the sample was sandwiched between 3-μm-thick films andanalyzed in transmission geometry. A beam-stop, short antiscatterextension, and antiscatter knife edge were used to minimize thebackground generated by air. Soller slits for the incident anddiffracted beams were used to minimize broadening and asymmetry fromaxial divergence. Diffraction patterns were collected using a scanningposition-sensitive detector (X'Celerator) located 240 mm from thespecimen and Data Collector software v. 2.2b or 5.5.

Example 4: Salt Data

Crystalline and anhydrous SRR G-1 besylate, camsylate, and napsylatesalts were successfully isolated. All three were obtained as 1:1stoichiometric salts. For these, seeding was crucial in providing highyields of crystalline salts that were not discolored. The XRPD patternsof the salts are compared with freebase Form A in FIG. 25. Scale-up andcharacterization of the salts are described in more detail in subsequentsections below.

SRR G-1 Besylate Form A

SRR G-1 Besylate Form A is an anhydrous 1:1 stoichiometric salt with anapparent melt onset near 186° C. Disproportionation of the salt in waterwas not evident.

The single-crystal structure of SRR G-1 Besylate Form A was determinedsuccessfully. Thus colorless needle having approximate dimensions of0.23×0.09×0.04 mm³, was mounted on a polymer loop in random orientation.Preliminary examination and data collection were performed on a RigakuSuperNova diffractometer, equipped with a copper anode microfocus sealedX-ray tube (Cu Kα λ=1.54184 Å) and a Dectris Pilatus3 R 200K hybridpixel array detector. Cell constants and an orientation matrix for datacollection were obtained from least-squares refinement using the settingangles of 13177 reflections in the range 4.2570°<θ<77.0580°. The spacegroup was determined by the program CRYSALISPRO to be P2₁. The data werecollected to a maximum diffraction angle (2θ) of 155.242° at roomtemperature.

Frames were integrated with CRYSALISPRO. A total of 26894 reflectionswere collected, of which 10520 were unique. Lorentz and polarizationcorrections were applied to the data. The linear absorption coefficientis 3.323 mm⁻¹ for Cu Kα radiation. An empirical absorption correctionusing CRYSALISPRO was applied. Transmission coefficients ranged from0.837 to 1.000. Intensities of equivalent reflections were averaged. Theagreement factor for the averaging was 3.3% based on intensity.

The structure was solved by direct methods using SHELXT. The remainingatoms were located in succeeding difference Fourier syntheses. Thestructure was refined using SHELXL-2014. Hydrogen atoms were refinedindependently. The structure was refined in full-matrix least-squares byminimizing the function:

Σw(|F _(o)|² −|F _(c)|²)²

where the weight, w, is defined as 1/[σ2(F_(o) ²)+(0.0401P)²+(0.3205P)],where P=(F_(o) ²+2F²)/3. Scattering factors were taken from the“International Tables for Crystallography”. Of the 10520 reflectionsused in the refinements, only the reflections with intensities largerthan twice their uncertainty [I>2σ(1)], 9411, were used in calculatingthe fit residual, R. The final cycle of refinement included 723 variableparameters, 1 restraint, and converged with respective unweighted andweighted agreement factors of:

R = ∑F_(o) − F_(c)/∑F_(o) = 0.0348$R_{w} = {\sqrt{\left( {\sum{{w\left( {F_{o}^{2} - F_{c}^{2}} \right)}^{2}/{\sum{w\left( F_{o}^{2} \right)}^{2}}}} \right)} = {0.0874}}$

The standard deviation of an observation of unit weight (goodness offit) was 1.05. The highest peak in the final difference Fourier had anelectron density of 0.311 e/↑³. The minimum negative peak had a value of−0.280 e/Å³.

The crystal system is monoclinic and the space group is P2₁. The cellparameters and calculated volume are: a=14.1207(3) Å, b=8.74139(11) Å,c=21.5361(4) Å, α=90°, β=106.1889(19°), γ=90°, V=2552.89(8) Å³. Theformula weight is 570.44 g mol⁻¹ with Z=4, resulting in a calculateddensity of 1.484 g cm⁻³. Further details of the crystal data andcrystallographic data collection parameters are summarized in Table 17.An atomic displacement ellipsoid drawing of Besylate Form A is shown inFIG. 26. The asymmetric unit shown contains two SRR G-1 cations and twobesylate anions. The —SO₃ moiety was modeled with rotational disorder onboth anions. A calculated XRPD pattern generated for Cu radiation usingMERCURY and the atomic coordinates, space group, and unit cellparameters from the single crystal structure is provided in FIG. 27 andcompared to the experimental pattern.

TABLE 17 SRR G-1 Besylate Form A Crystal Data and Data CollectionParameters Empirical formula C₂₇H₂₄BrNO₆S Formula weight (g mol⁻¹)570.44 Temperature (K) 299.64(13) Wavelength (Å) 1.54184 Crystal systemmonoclinic Space group P2₁ Unit cell parameters a = 14.1207(3) Å α = 90°b = 8.74139(11) Å β = 106.1889(19)° c = 21.5361(4) Å γ = 90° Unit cellvolume (Å³) 2552.89(8) Cell formula units, Z 4 Calculated density (gcm⁻³) 1.484 Absorption coefficient (mm⁻¹) 3.323 F(000) 1168 Crystal size(mm³) 0.23 × 0.09 × 0.04 Reflections used for cell measurement 13177 θrange for cell measurement 4.2570°-77.0580° Total reflections collected26894 Index ranges −17 ≤ h ≤ 17; 10 ≤ k ≤ 11; −25 ≤/≤ 27 θ range fordata collection θ_(min) = 3.259°, _(max) = 77.621° Completeness toθ_(max) 98.6% Completeness to θ_(full) = 67.684° 100% Absorptioncorrection multi-scan Transmission coefficient range 0.837-1.000Refinement method full matrix least-squares on F² Independentreflections 10520 [R_(int) = 0.0330, R_(σ) = 0.0387] Reflections [I >2σ(I)] 9411 Reflections/restraints/parameters 10520/1/723Goodness-of-fit on F² S = 1.05 Final residuals [I > 2σ(I)] R = 0.0348,R_(w) = 0.0874 Final residuals [all reflections] R = 0.0399, R_(w) =0.0904 Largest diff. peak and hole (e Å⁻³) 0.311, −0.280 Max/meanshift/standard uncertainty 0.001/0.000 Absolute structure determinationFlack parameter: −0.034(10)

The observed XRPD peaks for SRR G-1 Besylate Form A are listed in Table

TABLE 18 Diffraction angle 2θ (°) d-spacing (Å) Intensity (%)  4.26 ±0.20 20.737 ± 0.974  32  6.51 ± 0.20 13.567 ± 0.416  22  6.71 ± 0.2013.157 ± 0.392  100  8.54 ± 0.20 10.340 ± 0.242  14  8.72 ± 0.20 10.131± 0.232  7  9.18 ± 0.20 9.625 ± 0.209 10 10.97 ± 0.20 8.059 ± 0.147 1512.03 ± 0.20 7.351 ± 0.122 12 12.14 ± 0.20 7.283 ± 0.120 6 12.67 ± 0.206.980 ± 0.110 4 12.83 ± 0.20 6.895 ± 0.107 7 13.06 ± 0.20 6.774 ± 0.1034 13.25 ± 0.20 6.678 ± 0.100 15 13.45 ± 0.20 6.577 ± 0.097 13 13.67 ±0.20 6.471 ± 0.094 13 14.83 ± 0.20 5.970 ± 0.080 11 15.55 ± 0.20 5.694 ±0.073 4 15.95 ± 0.20 5.551 ± 0.069 13 16.14 ± 0.20 5.488 ± 0.068 6 16.24± 0.20 5.455 ± 0.067 5 16.36 ± 0.20 5.413 ± 0.066 7 16.86 ± 0.20 5.254 ±0.062 22 17.14 ± 0.20 5.169 ± 0.060 9 17.51 ± 0.20 5.060 ± 0.057 9 17.98± 0.20 4.930 ± 0.054 4 18.58 ± 0.20 4.771 ± 0.051 5 18.92 ± 0.20 4.687 ±0.049 20 19.09 ± 0.20 4.645 ± 0.048 7 19.42 ± 0.20 4.567 ± 0.047 1319.99 ± 0.20 4.437 ± 0.044 58 20.29 ± 0.20 4.373 ± 0.043 31 20.62 ± 0.204.304 ± 0.041 21 20.75 ± 0.20 4.277 ± 0.041 74 21.35 ± 0.20 4.158 ±0.038 25 21.40 ± 0.20 4.149 ± 0.038 31 21.46 ± 0.20 4.137 ± 0.038 3921.65 ± 0.20 4.102 ± 0.037 16 21.81 ± 0.20 4.072 ± 0.037 9 22.06 ± 0.204.026 ± 0.036 60 22.12 ± 0.20 4.015 ± 0.036 51 22.32 ± 0.20 3.980 ±0.035 28 22.45 ± 0.20 3.957 ± 0.035 15 22.67 ± 0.20 3.919 ± 0.034 423.01 ± 0.20 3.861 ± 0.033 8 23.14 ± 0.20 3.840 ± 0.033 7 23.55 ± 0.203.775 ± 0.032 14 23.70 ± 0.20 3.751 ± 0.031 13 23.78 ± 0.20 3.739 ±0.031 14 23.99 ± 0.20 3.706 ± 0.030 36 24.05 ± 0.20 3.697 ± 0.030 2624.18 ± 0.20 3.677 ± 0.030 15 24.36 ± 0.20 3.651 ± 0.030 15 24.43 ± 0.203.640 ± 0.029 19 24.79 ± 0.20 3.589 ± 0.029 3 25.23 ± 0.20 3.528 ± 0.0283 25.68 ± 0.20 3.466 ± 0.027 8 25.84 ± 0.20 3.446 ± 0.026 12 25.92 ±0.20 3.435 ± 0.026 21 26.30 ± 0.20 3.386 ± 0.025 11 26.69 ± 0.20 3.337 ±0.025 7 26.84 ± 0.20 3.319 ± 0.024 12 27.25 ± 0.20 3.270 ± 0.024 4 27.49± 0.20 3.242 ± 0.023 6 27.81 ± 0.20 3.205 ± 0.023 16 28.22 ± 0.20 3.160± 0.022 3 28.40 ± 0.20 3.140 ± 0.022 3 28.65 ± 0.20 3.114 ± 0.021 328.84 ± 0.20 3.093 ± 0.021 3 29.09 ± 0.20 3.068 ± 0.021 9 29.63 ± 0.203.012 ± 0.020 8 29.96 ± 0.20 2.980 ± 0.019 5 30.25 ± 0.20 2.952 ± 0.0196

The solution ¹H NMR spectrum is consistent with a 1:1 stoichiometricsalt of SRR G-1 and benzenesulfonic acid. Residual solvent is notevident, consistent with an unsolvated form.

Thermograms of SRR G-1 Besylate Form A are provided in FIG. 28.Negligible weight loss up to 186° C. is evident by TGA, consistent withan anhydrous form. The DSC exhibits a sharp endotherm with an onset near186° C. The event is likely due to a melt concurrent with decomposition.A small endotherm near 164° C. is also evident. The nature of thisendotherm is unknown.

The possibility of disproportionation in water was investigated. SRR G-1Besylate Form A was slurried in water for 4 days. The excess solids wererecovered and reanalyzed by XRPD for evidence of freebase orbenzenesulfonic acid. The recovered material was SRR G-1 Besylate FormA, indicating that disproportionation did not occur under the conditionevaluated.

The following describes a 1-gram scale procedure for generating SRR G-1Besylate Form A. A molar equivalent, 0.50 g, of benzenesulfonic acidmonohydrate was added to a vessel containing 1.17 g of SRR G-1 freebaseForm A. In addition, a small quantity of SRR G-1 Besylate Form A wasadded as seeds. Ethyl acetate, 7 mL, was added and followed bysonication. A predominant portion of the solids dissolved, resulting ina yellow solution, but was immediately followed by precipitation ofwhite solids. An additional 3 mL of ethyl acetate was added tofacilitate slurry transfer and filtration. The solids were recovered byvacuum filtration and rinsed with 4 mL of ethyl acetate followed byvacuum at room temperature overnight. Approximately 0.99 grams of SRRG-1 Besylate Form A was obtained.

SRR G-1 Camsylate Form A

SRR G-1 Camsylate Form A is an anhydrous 1:1 stoichiometric salt with anapparent melt onset of 172° C.

The XRPD pattern of SRR G-1 Camsylate Form A was successfully indexed,suggesting it is composed primarily of a single crystalline phase (FIG.29). SRR G-1 Camsylate Form A has a triclinic unit cell that canaccommodate two SRR G-1 cations and two camsylate anions. The formulaunit volume of 737.9 Å³ calculated from the indexing results would beconsistent with an anhydrous form with a calculated density of 1.451 gcm⁻³. The XRPD pattern also contains a small number of minor peaks thatare not associated with SRR G-1 Camsylate Form A, the known polymorphsof the freebase, or (+)-(1S)-camphor-10-sulfonic acid. These additionalpeaks are highlighted in FIG. 30 with asterisks.

The observed XRPD peaks for SRR G-1 Camsylate Form A are listed in Table19

TABLE 19 Diffraction angle 2θ (°) d-spacing (Å) Intensity (%)  5.97 ±0.20 14.786 ± 0.495  62  8.12 ± 0.20 10.876 ± 0.267  9  9.07 ± 0.209.738 ± 0.214 16 11.02 ± 0.20 8.025 ± 0.145 4 11.98 ± 0.20 7.379 ± 0.12332 12.69 ± 0.20 6.972 ± 0.109 30 13.41 ± 0.20 6.596 ± 0.098 39 14.53 ±0.20 6.089 ± 0.083 14 14.73 ± 0.20 6.009 ± 0.081 13 15.80 ± 0.20 5.604 ±0.070 14 16.23 ± 0.20 5.456 ± 0.067 26 17.79 ± 0.20 4.981 ± 0.056 3318.03 ± 0.20 4.916 ± 0.054 29 18.25 ± 0.20 4.858 ± 0.053 17 18.77 ± 0.204.724 ± 0.050 100 19.69 ± 0.20 4.506 ± 0.045 32 20.68 ± 0.20 4.292 ±0.041 18 21.28 ± 0.20 4.172 ± 0.039 12 21.62 ± 0.20 4.107 ± 0.038 921.81 ± 0.20 4.071 ± 0.037 14 22.13 ± 0.20 4.014 ± 0.036 14 22.33 ± 0.203.977 ± 0.035 19 22.54 ± 0.20 3.941 ± 0.035 14 22.70 ± 0.20 3.914 ±0.034 10 23.11 ± 0.20 3.845 ± 0.033 9 23.30 ± 0.20 3.815 ± 0.032 1723.45 ± 0.20 3.790 ± 0.032 20 23.86 ± 0.20 3.726 ± 0.031 10 24.12 ± 0.203.687 ± 0.030 3 24.57 ± 0.20 3.620 ± 0.029 8 25.12 ± 0.20 3.542 ± 0.0288 25.56 ± 0.20 3.482 ± 0.027 13 26.13 ± 0.20 3.408 ± 0.026 12 26.35 ±0.20 3.379 ± 0.025 16 26.78 ± 0.20 3.327 ± 0.024 8 27.22 ± 0.20 3.273 ±0.024 13 28.07 ± 0.20 3.176 ± 0.022 9 28.84 ± 0.20 3.093 ± 0.021 5 29.74± 0.20 3.002 ± 0.020 16

The solution ¹H NMR spectrum is consistent with a 1:1 stoichiometricsalt of SRR G-1 and (+)-(1S)-camphor-10-sulfonic acid. Residual solventis not evident, consistent with an unsolvated form.

Thermograms for SRR G-1 Camsylate Form A are provided in FIG. 31.Negligible weight loss up to 171° C. is evident by TGA, consistent withan anhydrous form. The DSC exhibits a sharp endotherm with an onset near172° C. The event is likely due to a melt concurrent with decomposition.

The following describes a 750-mg scale procedure for generating SRR G-1Camsylate Form A. Less than a molar equivalent (0.9), 0.43 g, of(+)-(1S)-camphor-10-sulfonic acid was added to a vessel containing asuspension composed of 0.86 g of SRR G-1 freebase Form A and 10 mL ofethyl acetate, providing a yellow suspension with a small amount ofundissolved solids. Seeds of SRR G-1 Camsylate Form A were added and thesuspension was sonicated causing immediate precipitation. The sample wassonicated for an additional ˜10 minutes and then left to slurry forapproximately 1 hour. The white solids were recovered by vacuumfiltration and rinsed with 2 mL of ethyl acetate followed by vacuum atroom temperature overnight. Approximately 0.75 grams of SRR G-1Camsylate Form A was obtained.

SRR G-1 Napsylate Form A

SRR G-1 Napsylate Form A is an anhydrous 1:1 stoichiometric salt with anapparent melt onset of 194° C. Based on XRPD results from an aqueousslurry, disproportionation of the salt does occur in water.

The XRPD pattern of SRR G-1 Napsylate Form A was successfully indexed,suggesting it is composed primarily of a single crystalline phase (FIG.32). SRR G-1 Napsylate Form A has monoclinic unit cell that canaccommodate four SRR G-1 cations and four napsylate anions. The formulaunit volume of 707.3 Å³ calculated from the indexing results would beconsistent with an anhydrous form with a calculated density of 1.457 gcm⁻³. The XRPD pattern also contains a small, weak peak near 4.4° (20)that is not associated with SRR G-1 Napsylate Form A, the knownpolymorphs of the freebase, or napthtlane-2-sulfonic acid.

The observed XRPD peaks for SRR G-1 Napsylate Form A are listed in Table20

TABLE 20 Diffraction angle 2θ (°) d-spacing (Å) Intensity (%)  6.17 ±0.20 14.321 ± 0.464  88  8.91 ± 0.20 9.913 ± 0.222 26 10.16 ± 0.20 8.703± 0.171 10 10.40 ± 0.20 8.496 ± 0.163 17 11.68 ± 0.20 7.570 ± 0.129 1112.38 ± 0.20 7.145 ± 0.115 17 12.63 ± 0.20 7.001 ± 0.110 68 12.84 ± 0.206.891 ± 0.107 64 13.18 ± 0.20 6.713 ± 0.101 23 13.75 ± 0.20 6.433 ±0.093 31 14.39 ± 0.20 6.151 ± 0.085 54 15.01 ± 0.20 5.897 ± 0.078 2115.26 ± 0.20 5.800 ± 0.076 13 16.15 ± 0.20 5.485 ± 0.067 23 16.79 ± 0.205.277 ± 0.062 73 17.07 ± 0.20 5.189 ± 0.060 48 17.21 ± 0.20 5.148 ±0.059 14 17.64 ± 0.20 5.025 ± 0.057 95 17.90 ± 0.20 4.950 ± 0.055 1218.23 ± 0.20 4.863 ± 0.053 10 18.62 ± 0.20 4.762 ± 0.051 21 18.97 ± 0.204.674 ± 0.049 22 19.22 ± 0.20 4.615 ± 0.048 100 19.44 ± 0.20 4.563 ±0.046 53 19.67 ± 0.20 4.510 ± 0.045 11 20.06 ± 0.20 4.422 ± 0.044 1820.43 ± 0.20 4.344 ± 0.042 35 20.76 ± 0.20 4.274 ± 0.041 22 21.13 ± 0.204.201 ± 0.039 20 21.26 ± 0.20 4.175 ± 0.039 46 21.78 ± 0.20 4.077 ±0.037 48 21.91 ± 0.20 4.053 ± 0.037 30 22.11 ± 0.20 4.017 ± 0.036 2622.60 ± 0.20 3.931 ± 0.034 65 23.14 ± 0.20 3.841 ± 0.033 18 23.38 ± 0.203.802 ± 0.032 56 23.63 ± 0.20 3.762 ± 0.031 14 24.40 ± 0.20 3.645 ±0.029 24 24.60 ± 0.20 3.615 ± 0.029 9 25.13 ± 0.20 3.541 ± 0.028 1425.30 ± 0.20 3.517 ± 0.027 19 25.47 ± 0.20 3.495 ± 0.027 23 25.85 ± 0.203.444 ± 0.026 19 26.07 ± 0.20 3.415 ± 0.026 63 26.60 ± 0.20 3.348 ±0.025 18 26.88 ± 0.20 3.315 ± 0.024 7 27.38 ± 0.20 3.254 ± 0.023 1527.63 ± 0.20 3.226 ± 0.023 50 28.27 ± 0.20 3.154 ± 0.022 9 28.67 ± 0.203.111 ± 0.021 11 28.90 ± 0.20 3.087 ± 0.021 9 29.02 ± 0.20 3.075 ± 0.02112 29.15 ± 0.20 3.061 ± 0.021 17

The solution ¹H NMR spectrum is consistent with a 1:1 stoichiometricsalt of SRR G-1 and naphthalene-2-sulfonic acid. Residual solvent is notevident, consistent with an unsolvated form.

Thermograms for SRR G-1 Napsylate Form A are provided in FIG. 33.Approximately 0.5% weight loss up to 193° C. is evident by TGA. Themajority of the loss occurs above ˜100° C. Because organic solvent wasnot observed by NMR, discussed above, it is assumed the loss is due tothe volatilization of approximately 0.2 mol/mol water. This suggeststhat the salt may exhibit limited hygroscopicity. The DSC exhibits asharp endotherm with an onset near 194° C. The event is likely due to amelt concurrent with decomposition.

The possibility of disproportionation in water was investigated. SRR G-1Napsylate Form A was slurried in water for 5 days. The excess solidswere recovered and reanalyzed by XRPD for evidence of freebase ornaphthalene-2-sulfonic acid. The recovered material was freebase Form A,indicating that disproportionation occurred under the conditionevaluated.

The following describes a 600-mg scale procedure for generating SRR G-1Napsylate Form A. Seeds of SRR G-1 Napsylate Form A and a molarequivalent, 0.39 g, of naphthalene-2-sulfonic acid was added to a vesselcontaining a suspension of 0.73 g of SRR G-1 freebase Form A and 9 mL ofethyl acetate. The yellow suspension with a small amount of undissolvedsolids was sonicated and a white precipitation occurred. The slurry wassonicated for an additional ˜5 minutes and the solids were thenrecovered by vacuum filtration, rinsed with 2 mL of ethyl acetate, anddried under vacuum at room temperature overnight. Approximately 0.63grams of SRR G-1 Napsylate Form A was obtained.

Example 5: YUMM1.7 Proliferation Assay

YUMM1.7 cells were cultured for at least 1 passage after thawing andwere cultured in DMEM with 5% FBS (Invitrogen) and 1%antibiotic-antimycotic (gibco) at 37 C 5% CO₂. Proliferation assays wereperformed by plating 15,000 cells in 12-well plates with 5 replicatesper condition tested. Media and drugs were refreshed on Day 2. On Day 4,cells were trypsinized using 0.25 ml 0.05% Trypsin with EDTA(Invitrogen) for 5 minutes to detach from the plate, mixed with 0.75 mlof culture media, and counted using a hemocytometer.

The average cell count after 4 days of growth in a YUMM1.7 proliferationassay conducted with 500 nM of each composition are shown in Table 21.The same data is shown in graphical form in FIG. 34. Cell counts are inthe tens of thousands (i.e. 100 is about 1,000,000). Starting cellnumbers were 15,000. RSS G-1 had approximately the same number ofdoublings as the vehicle. Racemic G-1 reduced doubling to about half ofthat seen with the vehicle. Surprisingly, SRR G-1 reduced doubling toless than 1/10^(th) of that seen with the vehicle rather than just ¼ aswould be expected from the reduction caused by G-1.

TABLE 21 Vehicle Racemic G-1 SRR G-1 RSS G-1 104 16 2 83 84 18 3 94 10819 2 79 117 17 3 111 97 24 2 107 Averages 102 18.8 2.4 94.8 Doublings8.2 3.4 0.6 7.9

Example 6: Preclinical Rat Pharmacokinetic Results

Plasma concentrations of SRR G-1 free base, SRR G-1 besylate, and SRRG-1 napsylate in rats was determined after oral dosing. Three fasted,male rats were treated with 10 mg/kg SRR G-1 free base, SRR G-1besylate, or SRR G-1 napsylate delivered orally as a suspension in 0.5%hydroxypropyl methylcellulose, 99.5% water. Plasma was isolated at 0.5,1, 2, 4, 8, and 24 hours after SRR G-1 administration, and plasmaconcentrations were determined using LC-MS/MS. The results are shown inTables 22-24 for each respectively. Graphical representation of thisdata is shown in FIGS. 35-37. FIG. 38 shows a comparison of all threeresults.

TABLE 22 SRR G-1 Free Base LLOQ 0.100 ng/mL Plasma PO (10 mg/kg) ULOQ300 ng/mL Conc. (ng/mL) Time (h) R1 R2 R3 0.500 1.91 2.60 1.23 1.00 3.966.03 3.15 2.00 2.96 7.56 2.71 4.00 2.18 4.67 1.39 8.00 1.72 1.58 1.4724.0 0.127 0.169 BQL

TABLE 23 SRR G-1 Besylate LLOQ 0.100 ng/mL Plasma PO (10 mg/kg) ULOQ 300ng/mL Conc. (ng/mL) Time (h) R4 R5 R6 0.500 39.2 35.2 34.1 1.00 43.942.0 44.5 2.00 28.5 30.6 32.3 4.00 20.0 16.7 19.5 8.00 8.17 5.78 6.1824.0 0.837 0.668 0.647

TABLE 24 SRR G-1 Napsylate LLOQ 0.100 ng/mL Plasma PO (10 mg/kg) ULOQ300 ng/mL Conc. (ng/mL) Time (h) R7 R8 R9 0.500 62.2 50.6 22.6 1.00 81.967.5 33.2 2.00 46.6 39.9 24.3 4.00 14.6 14.0 15.6 8.00 5.45 6.09 3.9724.0 0.641 0.558 0.435

Example 7: ADME Toxicology of SRR G-1 and RSS G-1

ADME-Tox: In Vitro Absorption

Drug Transporter (Fluorometric Inhibition)

The percent of control was calculated using the following equation. Thepercent of inhibition was calculated by subtracting the percent ofcontrol from 100. The IC50 value (concentration causing a half-maximalinhibition of the control value) was determined by non-linear regressionanalysis of the concentration-response curve using the Hill equation.

${{Control}\mspace{14mu}(\%)} = {\frac{{Compound} - {Background}}{{T\; 1} - {Background}}*100}$

Compound is the individual reading in the presence of the test compound.T1 is the mean reading in the absence of the test compound. Background(for P-gp and BCRP) is the mean reading in the presence of the highesteffective concentration of the reference inhibitor. Background (forOATP1B1, OATP1B3, OAT1, OAT3, and OCT2) is the mean reading in theabsence of both the test compound and the substrate.

ADME-Tox: In Vitro Metabolism

Cytochrome P450 Inhibition (HPLC-UV/VIS and HPLC-MS/MS detection)

Peak areas corresponding to the metabolite of each substrate wererecorded. The percent of control activity was then calculated bycomparing the peak area obtained in the presence of the test compound tothat obtained in the absence of the test compound. Subsequently, thepercent inhibition was calculated by subtracting the percent controlactivity from 100 for each compound. IC50 values (concentration causinga half-maximal inhibition of control values) were determined bynon-linear regression analysis of the concentration-response curve usingHill equation curve fitting.

Transporter inhibition results—When assayed with 10 μM of SRR G-1 or RSSG-1, the following transporters were inhibited more than 50%. For SRRG-1: OATP1B1 82.5%. For RSS G-1: OCT2—53.2%, OATP1B1—91.2%, andOATP1B3—74.3%.

Cytochrome P450 inhibition results—When assayed with 10 μM of SRR G-1 orRSS G-1, the following were inhibited more than 50%. For SRR G-1:CYP2D6—74.3% and CYP2C8—66.7%. For RSS G-1: CYP2C9-50.4%.

Cytochrome P450 induction results—Hepatocytes from three different humancell lines were incubated with SRR G-1 or RSS G-1 at 1 μM, 10 μM, and100 μM. For SRR G-1: CYP1A2 was induced at both 1 μM and 10 μM in only 1of the 3 cell lines and CYP3A4 was induced at only 10 μM in 2 of the 3cell lines. For RSS G-1: CYP1A2 was induced at both 10 μM and 100 μM in2 of the 3 cell lines.

Example 8: Off Target Selectivity Assays

GPCR cAMP Modulation

Cell Handling—cAMP Hunter cell lines were expanded from freezer stocksaccording to standard procedures. Cells were seeded in a total volume of20 μm into white walled, 384-well microplates and incubated at 37° C.for the appropriate time prior to testing. cAMP modulation wasdetermined using the DiscoverX HitHunter cAMP XS+ assay.

Gs Agonist Format—For agonist determination, cells were incubated withsample to induce response. Media was aspirated from cells and replacedwith 15 μL 2:1 HBSS/10 mM Hepes: cAMP XS+ Ab reagent. Intermediatedilution of sample stocks was performed to generate 4× sample in assaybuffer. 4.5 μL of 4× sample was added to cells and incubated at 37° C.or room temperature for 30 or 60 minutes. Final assay vehicleconcentration was 1%.

Gi Agonist Format—For agonist determination, cells were incubated withsample in the presence of EC₈₀ forskolin to induce response. Media wasaspirated from cells and replaced with 15 μL 2:1 HBSS/10 mM Hepes:cAMPXS+ Ab reagent. Intermediate dilution of sample stocks was performed togenerate 4× sample in assay buffer containing 4× EC₈₀ forskolin. 4.5 μLof 4× sample was added to cells and incubated at 37° C. or roomtemperature for 30 or 60 minutes. Final assay vehicle concentration was1%.

Antagonist Format—For antagonist determination, cells were pre-incubatedwith sample followed by agonist challenge at the ECso concentration.Media was aspirated from cells and replaced with 10 μL 1:1 HBSS/Hepes:cAMP XS+Ab reagent. 5 μL of 4× compound was added to the cells andincubated at 37° C. or room temperature for 30 minutes. 4.5 μL of 4×EC₈₀ agonist was added to cells and incubated at 37° C. or roomtemperature for 30 or 60 minutes. For Gi coupled GPCRs, EC₈₀ forskolinwas included.

Signal Detection—After appropriate compound incubation, assay signal wasgenerated through incubation with 20 μL cAMP XS+ED/CL lysis cocktail forone hour followed by incubation with 20 μL cAMP XS+EA reagent for threehours at room temperature. Microplates were read following signalgeneration with a PerkinElmer Envision™ instrument for chemiluminescentsignal detection.

Data Analysis—Compound activity was analyzed using CBIS data analysissuite (ChemInnovation, CA). For Gs agonist mode assays, percentageactivity was calculated using the following formula: %Activity=100%×(mean RLU of test sample−mean RLU of vehiclecontrol)/(mean RLU of MAX control−mean RLU of vehicle control). For Gsantagonist mode assays, percentage inhibition was calculated using thefollowing formula: % Inhibition=100%×(1−(mean RLU of test sample−meanRLU of vehicle control)/(mean RLU of EC₈₀ control−mean RLU of vehiclecontrol)). For Gi agonist mode assays, percentage activity wascalculated using the following formula: % Activity=100%×(1−(mean RLU oftest sample−mean RLU of MAX control)/(mean RLU of vehicle control−meanRLU of MAX control)). For Gi antagonist or negative allosteric modeassays, percentage inhibition was calculated using the followingformula: % Inhibition=100%×(mean RLU of test sample−mean RLU of EC₈₀control)/(mean RLU of forskolin positive control−mean RLU of EC₈₀control). For Primary screens, percent response was capped at 0% or 100%where calculated percent response returned a negative value or a valuegreater than 100, respectively.

Calcium Mobilization

Cell Handling—Cell lines were expanded from freezer stocks according tostandard procedures. Cells (10,000 cells/well) were seeded in a totalvolume of 50 μL (200 cells/_L) into black-walled, clear-bottom,Poly-D-lysine coated 384-well microplates and incubated at 37° C. forthe appropriate time prior to testing. DMSO concentration for allreadouts is ≤0.2%.

Dye Loading—Assays were performed in 1× Dye Loading Buffer consisting of1× Dye (DiscoverX, Calcium No WashPLUS kit, Catalog No. 90-0091), 1×Additive A and 2.5 mM Probenecid in HBSS/20 mM Hepes. Probenicid wasprepared fresh. Cells were loaded with dye prior to testing. Media wasaspirated from cells and replaced with 25 μL Dye Loading Buffer. Cellswere incubated for 45 minutes at 37° C. and then 20 minutes at roomtemperature.

Agonist Format—For agonist determination, cells were incubated withsample to induce response. After dye loading, cells were removed fromthe incubator and 25 μL of 2× compound in HBSS/20 mM Hepes was addedusing a FLIPR Tetra (MDS). Compound was added and agonist activity wasmeasured on a FLIPR Tetra. Calcium mobilization was monitored for 2minutes with a 5 second baseline read.

Antagonist Format—Cells were preincubated with sample, dye loaded, movedto the FLIPR Tetra (MDS) and then challenged with an agonist at the EC80concentration. Calcium mobilization was monitored for 2 minutes with a 5second baseline read.

Data Analysis—FLIPR read—Area under the curve was calculated for theentire two minute read. Compound activity was analyzed using CBIS dataanalysis suite (ChemInnovation, CA). For agonist mode assays, percentageactivity was calculated using the following formula: %Activity=100%×(mean RFU of test sample−mean RFU of vehiclecontrol)/(mean MAX RFU control ligand−mean RFU of vehicle control). Forantagonist mode assays, percentage inhibition was calculated using thefollowing formula: % Inhibition=100%×(1−(mean RFU of test sample−meanRFU of vehicle control)/(mean RFU of EC₈₀ control−mean RFU of vehiclecontrol)). For Primary screens, percent response was capped at 0% or100% where calculated percent response returned a negative value or avalue greater than 100, respectively.

Nuclear Hormone Receptor

Cell Handling—PathHunter NHR cell lines were expanded from freezerstocks according to standard procedures. Cells were seeded in a totalvolume of 20 μL into white walled, 384-well microplates and incubated at37° C. for the appropriate time prior to testing. Assay media containedcharcoal-dextran filtered serum to reduce the level of hormones present.

Agonist Format—For agonist determination, cells were incubated withsample to induce response. Intermediate dilution of sample stocks wasperformed to generate 5× sample in assay buffer. 3.5 μL of 5× sample wasadded to cells and incubated at 37° C. or room temperature for 3-16hours. Final assay vehicle concentration was 1%.

Antagonist Format—For antagonist determination, cells were pre-incubatedwith antagonist followed by agonist challenge at the EC₈₀ concentration.Intermediate dilution of sample stocks was performed to generate 5×sample in assay buffer. 3.5 μL of 5× sample was added to cells andincubated at 37° C. or room temperature for 60 minutes. Vehicleconcentration was 1%. 4.5 μL of 6× EC₈₀ agonist in assay buffer wasadded to the cells and incubated at 37° C. or room temperature for 3-16hours.

Signal Detection—Assay signal was generated through a single addition of12.5 or 15 μL (50% v/v) of PathHunter Detection reagent cocktail,followed by a one hour incubation at room temperature. Microplates wereread following signal generation with a PerkinElmer Envision™ instrumentfor chemiluminescent signal detection.

Data Analysis—Compound activity was analyzed using CBIS data analysissuite (ChemInnovation, CA). For agonist mode assays, percentage activitywas calculated using the following formula: % Activity=100%×(mean RLU oftest sample−mean RLU of vehicle control)/(mean MAX control ligand−meanRLU of vehicle control). For antagonist mode assays, percentageinhibition was calculated using the following formula: %Inhibition=100%×(1−(mean RLU of test sample−mean RLU of vehiclecontrol)/(mean RLU of EC₈₀ control−mean RLU of vehicle control)). Notethat for select assays, the ligand response produces a decrease inreceptor activity (inverse agonist with a constitutively active target).For those assays inverse agonist activity was calculated using thefollowing formula: % Inverse Agonist Activity=100%×((mean RLU of vehiclecontrol−mean RLU of test sample)/(mean RLU of vehicle control−mean RLUof MAX control)). For Primary screens, percent response was capped at 0%or 100% where calculated percent response returned a negative value or avalue greater than 100, respectively.

KINOMEscan Binding Assays

Protein Expression—For most assays, kinase-tagged T7 phage strains weregrown in parallel in 24-well blocks in an E. coli host derived from theBL21 strain. E. coli were grown to log-phase and infected with T7 phagefrom a frozen stock (multiplicity of infection=0.4) and incubated withshaking at 32° C. until lysis (90-150 minutes). The lysates werecentrifuged (6,000×g) and filtered (0.2 μm) to remove cell debris. Theremaining kinases were produced in HEK-293 cells and subsequently taggedwith DNA for qPCR detection.

Capture Ligand Production—Streptavidin-coated magnetic beads weretreated with biotinylated small molecule ligands for 30 minutes at roomtemperature to generate affinity resins for kinase assays. The ligandedbeads were blocked with excess biotin and washed with blocking buffer(SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unboundligand and to reduce non-specific phage binding.

Binding Reaction Assembly—Binding reactions were assembled by combiningkinases, liganded affinity beads, and test compounds in 1× bindingbuffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20, 6 mM DTT). All reactionswere performed in polypropylene 384-well plates in a final volume of0.02 mL. The assay plates were incubated at room temperature withshaking for 1 hour and the affinity beads were washed with wash buffer(lx PBS, 0.05% Tween 20). The beads were then re-suspended in elutionbuffer (1×PBS, 0.05% Tween 20, 0.5 μM non-biotinylated affinity ligand)and incubated at room temperature with shaking for 30 minutes. Thekinase concentration in the eluates was measured by qPCR.

Signal Detection—The kinase concentration in the eluates was measured byqPCR. qPCR reactions were assembled by adding 2.5 μL of kinase eluate to7.5 μL of qPCR master mix containing 0.15 μM amplicon primers and 0.15μM amplicon probe. The qPCR protocol consisted of a 10 minute hot startat 95° C., followed by 35 cycles of 95° C. for 15 seconds, 60° C. for 1minute.

Data Analysis—Percent Response Calculation

$\left( \frac{{{test}\mspace{14mu}{compound}\mspace{14mu}{signal}} - {{positive}\mspace{14mu}{control}\mspace{14mu}{signal}}}{{{negative}\mspace{14mu}{compound}\mspace{14mu}{signal}} - {{positive}\mspace{14mu}{control}\mspace{14mu}{signal}}} \right) \times 100$

Test Compound=SRR G-1

Negative control=DMSO (100% Ctrl)Positive control=control compound (0% Ctrl)Percent of Control was converted to Percent Response using formula:Percent Response=(100−Percent Control). For Primary screens, percentresponse was capped at 0% or 100% where calculated percent responsereturned a negative value or a value greater than 100, respectively.

Data Analysis—Binding Constants (Kds)

Binding constants (Kds) were calculated with a standard doseresponsecurve using the Hill equation:

${Response} = {{Background} + \left( \frac{{Signal} - {Background}}{1 + \left( {{Kd}^{{Hill}\mspace{11mu}{Slope}}/{Dose}^{{Hill}\mspace{11mu}{Slope}}} \right)} \right)}$

The Hill Slope was set to −1.

Curves were fitted using a non-linear least square fit with theLevenberg-Marquardt algorithm.

Ion Channel Assays

Cell Handling—Cell lines were expanded from freezer stocks according tostandard procedures. Cells were seeded in a total volume of 20 μL intoblack-walled, clear-bottom, Poly-D-lysine coated 384-well microplatesand incubated at 37° C. for the appropriate time prior to testing.

Dye Loading—Assays were performed in 1×Dye Loading Buffer consisting of1× Dye, and 2.5 mM Probenecid when applicable. Probenicid was preparedfresh. Cells were loaded with dye prior to testing. Cells were incubatedfor 30-60 minutes at 37° C.

Agonist/Opener Format—For agonist determination, cells were incubatedwith sample to induce response. Intermediate dilution of sample stockswas performed to generate 2-5× sample in assay buffer. 10-25 μL of 2-5×sample was added to cells and incubated at 37° C. or room temperaturefor 30 minutes. Final assay vehicle concentration was 1%.

Antagonist/Blocker Format—For antagonist determination, cells werepre-incubated with sample. Intermediate dilution of sample stocks wasperformed to generate 2-5× sample in assay buffer. After dye loading,cells were removed from the incubator and 10-25 μL 2-5× sample was addedto cells in the presence of EC₈₀ agonist when appropriate. Cells wereincubated for 30 minutes at room temperature in the dark to equilibrateplate temperature. Vehicle concentration was 1%.

Signal Detection—Compound activity was measured on a FLIPR Tetra (MDS).

Data Analysis—Compound activity was analyzed using CBIS data analysissuite (ChemInnovation, CA). For agonist mode assays, percentage activitywas calculated using the following formula: the following formula: %Activity=100%×(mean RLU of test sample−mean RLU of vehiclecontrol)/(mean MAX control ligand−mean RLU of vehicle control). Forantagonist percentage inhibition was calculated using the followingformula: % Inhibition=100%×(1−(mean RLU of test sample−mean RLU ofvehicle control)/(mean RLU of EC₈₀ control−mean RLU of vehiclecontrol)). For Primary screens, percent response was capped at 0% or100% where calculated percent response returned a negative value or avalue greater than 100, respectively.

Transporter Assays

Cell Handling—Cell lines were expanded from freezer stocks according tostandard procedures. Cells were seeded in a total volume of 25 μL intoblack-walled, clear-bottom, Poly-D-lysine coated 384-well microplatesand incubated at 37° C. for the appropriate time prior to testing.

Blocker/Antagonist Format—After cell plating and incubation, media wasremoved and 25 μL of 1× compound in 1×HBSS/0.1% BAS was added. Compoundswere incubated with cells at 37° C. for 30 minutes.

Dye Loading—Assays were performed in 1×Dye Loading Buffer consisting of1× Dye, 1×HBSS/20 mM Hepes. After compound incubation, 25 μL of 1× dyewas added to wells. Cells were incubated for 30-60 minutes at 37° C.

Signal Detection—After dye incubation, microplates were transferred to aPerkinElmer Envision™ instrument for fluorescence signal detection.

Data Analysis—Compound activity was analyzed using CBIS data analysissuite (ChemInnovation, CA). For blocker mode assays, percentageinhibition was calculated using the following formula: %Inhibition=100%×(1−(mean RLU of test sample−mean RLU of vehiclecontrol)/(mean RLU of positive control−mean RLU of vehicle control)).For Primary screens, percent response was capped at 0% or 100% wherecalculated percent response returned a negative value or a value greaterthan 100, respectively.

Enzymatic Assays

Enzyme Preparations—Enzyme preparations were sourced from variousvendors-AChE (R&D Systems), COX1 and COX2 (BPS Bioscience), MAOA(Sigma), PDE3A and PDE4D2 (Signal Chem).

Enzyme Activity Assays—Enzymatic assays determine the enzymatic activityby measuring either the consumption of substrate or production ofproduct over time. Different detection methods were used in eachenzymatic assay to measure the concentrations of substrates andproducts. AChE: Enzyme and test compound were preincubated for 15minutes at room temp before substrate addition. Acetylthiocholine andDTNB were added and incubated at room temperature for 30 minutes. Signalwas detected by measuring absorbance at 405 nm. COX1 & COX2: Enzymestocks were diluted in Assay Buffer (40 mM Tris-HCl, 1×PBS, 0.5 mMPhenol, 0.01% Tween-20+100 nM Hematin) and allowed to equilibrate withcompounds at room temperature for 30 minutes (binding incubation).Arachidonic acid (1.7 μM) and Ampliflu Red (2.5 μM) were prepared anddispensed into a reaction plate. Plates were read immediately on afluorimeter with the emission detection at 590 nm and excitationwavelength 544 nm. MAOA: Enzyme and test compound were preincubated for15 minutes at 37° C. before substrate addition. The reaction wasinitiated by addition of kynuramine and incubated at 37° C. for 30minutes. The reaction was terminated by addition of NaOH. The amount of4-hydroquioline formed was determined through spectrofluorimetricreadout with the emission detection at 380 nm and excitation wavelength310 nm. PDE3A & PDE4D2: Enzyme and test compound were preincubated for15 minutes at room temp before substrate addition. cAMP substrate (at aconcentration equal to EC₈₀) was added and incubated at room temperaturefor 30 minutes. Enzyme reaction was terminated by addition of 9 mM IBMX.Signal was detected using the HitHunter® cAMP detection kit.

Signal Detection—For each assay, microplates were transferred to aPerkinElmer Envision™ instrument and readout as described.

Data Analysis—Compound activity was analyzed using CBIS data analysissuite (ChemInnovation, CA). For enzyme activity assays, percentageinhibition was calculated using the following formula: %Inhibition=100%×(1−(mean RLU of test sample−mean RLU of vehiclecontrol)/(mean RLU of positive control−mean RLU of vehicle control)).For Primary screens, percent response was capped at 0% or 100% wherecalculated percent response returned a negative value or a value greaterthan 100, respectively.

Results of the Off Target Selectivity Assays

Both SRR G-1 and RSS G-1 were tested for selectivity against potentialoff targets in 78 assays in a dose response format at concentrations upto 1004. SRR G-1 only had a measurable IC₅₀ or EC₅₀ on Cannabinoidreceptor 1 at 2.5 μM, HTR2B at 8.2 μM, OPRD1 at 0.87 μM, and OPRM1 at6.68 μM. RSS G-1 only had a measurable IC₅₀ or EC₅₀ on Cannabinoidreceptor 1 at 3.1 μM, ADRA2A at 2.07 μM, HTR1A at 2.1 μM, and AR at 4.76μM.

1. A method of treating a disease or disorder in a subject in needthereof, comprising: a) administering to the subject a therapeuticallyeffective amount of a compound of the formula:

1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-oneSRR G-1 or a pharmaceutically acceptable salt thereof, wherein thecompound comprises a crystalline Form A characterized by an XRPD patternhaving peaks expressed in degrees 2θ (±0.20) at about 5.75, about 20.54,about 20.71, about 21.25, and about 21.86, wherein the crystalline FormA has a melt onset of about 176° C., wherein the crystalline Form A isanhydrous, wherein the crystalline Form A exhibits low hygroscopicity,and wherein the crystalline Form A exhibits a calculated density ofabout 1.485 g cm⁻³, and b) alleviating one or more symptoms associatedwith the disease or disorder in the subject.
 2. The method of claim 1,wherein the compound is a free base.
 3. The method of claim 1, whereinactivation of G-protein coupled estrogen receptor (GPER) by the compoundalleviates the one or more symptoms associated with the disease ordisorder.
 4. The method of claim 1, wherein cells causing or involved inthe disease or disorder express G-protein coupled estrogen receptor(GPER).
 5. The method of claim 1 further comprising administering to thesubject one or more additional therapeutic agents selected from thegroup consisting of an immunotherapy agent, a chemotherapy agent, atargeted kinase inhibitor, a histone deacetylase inhibitor, abromodomain inhibitor, and combinations thereof.
 6. The method of claim1, wherein the subject is a human or an animal.
 7. The method of claim1, wherein the disease or disorder is selected from the group consistingof cancer, endometritis, prostatitis, polycystic ovarian syndrome,urinary incontinence, hormone-related disorders, hearing disorders, hotflashes, profuse sweating, hypertension, stroke, ischemia, myocardialinfarction, dilated cardiomyopathy, obesity, insulin resistance,osteoporosis, atherosclerosis, symptoms of menopause, inflammation,rheumatoid arthritis, osteoarthritis, lymphoproliferative disorders,myeloproliferative disorders, eosinophilia, histiocytosis, paroxysmalnocturnal hemoglobinuria, systemic mastocytosis, venous thrombosis,embolisms, depression, insomnia, anxiety, neuropathy, multiplesclerosis, Parkinson's disease, Alzheimer's disease, inflammatory boweldisease, Crohn's disease, celiac disease, proteinuric renal disease,vascular disease, and thymic atrophy.
 8. The method of claim 7, whereinthe disease or disorder is cancer.
 9. The method of claim 8, wherein thecancer is selected from the group consisting of reproductive cancers,hormone-dependent cancers, leukemia, colorectal cancer, prostate cancer,breast cancer, ovarian carcinoma, endometrial cancer, uterinecarcinosarcoma, stomach cancer, rectal cancer, liver cancer, pancreaticcancer, lung cancer, uterine cancer, cervical cancer, cervix utericancer, corpus uteri cancer, ovary cancer, testicular cancer, bladdercancer, renal cancer, brain/CNS cancer, head and neck cancer, throatcancer, Hodgkin's disease, non-Hodgkin's lymphoma, B-cell lymphoma,T-cell lymphoma, uveal melanoma, triple negative breast cancer, multiplemyeloma, melanoma, acute leukemia, lymphocytic leukemia, hairy cellleukemia, acute myelogenous leukemia, Ewing's sarcoma, small cell lungcancer, non-small cell lung cancer, choriocarcinoma, rhabdomyosarcoma,Wilms's Tumor, neuroblastoma, cancer of the mouth/pharynx, cancer of theesophagus, cancer of the larynx, kidney cancer, lymphoma, Burkittlymphoma, sarcoma, angiosarcoma, glioblastoma, medulloblastoma,astrocytoma, and Merkel cell carcinoma.
 10. A method of treating type 2diabetes in a subject in need thereof, comprising: a) administering tothe subject a therapeutically effective amount of a compound of theformula:

1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-oneSRR G-1 or a pharmaceutically acceptable salt thereof, wherein thecompound comprises a crystalline Form A characterized by an XRPD patternhaving peaks expressed in degrees 2θ (±0.20) at about 5.75, about 20.54,about 20.71, about 21.25, and about 21.86, wherein the crystalline FormA has a melt onset of about 176° C., wherein the crystalline Form A isanhydrous, wherein the crystalline Form A exhibits low hygroscopicity,and wherein the crystalline Form A exhibits a calculated density ofabout 1.485 g cm⁻³; and b) alleviating one or more symptoms of type 2diabetes in the subject.
 11. A method of increasing or preventing lossof skin pigmentation in a subject in need thereof, comprising: a)administering to the subject a therapeutically effective amount of acompound of the formula:

1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-oneSRR G-1 or a pharmaceutically acceptable salt thereof, wherein thecompound comprises a crystalline Form A characterized by an XRPD patternhaving peaks expressed in degrees 2θ (±0.20) at about 5.75, about 20.54,about 20.71, about 21.25, and about 21.86, wherein the crystalline FormA has a melt onset of about 176° C., wherein the crystalline Form A isanhydrous, wherein the crystalline Form A exhibits low hygroscopicity,and wherein the crystalline Form A exhibits a calculated density ofabout 1.485 g cm⁻³; and increasing or preventing loss of skinpigmentation in the subject.
 12. A method of skin protection in asubject in need thereof comprising: a) administering to a subject atherapeutically effective amount of a compound of the formula:

1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-oneSRR G-1 or a pharmaceutically acceptable salt thereof, wherein thecompound comprises a crystalline Form A characterized by an XRPD patternhaving peaks expressed in degrees 2θ (±0.20) at about 5.75, about 20.54,about 20.71, about 21.25, and about 21.86, wherein the crystalline FormA has a melt onset of about 176° C., wherein the crystalline Form A isanhydrous, wherein the crystalline Form A exhibits low hygroscopicity,and wherein the crystalline Form A exhibits a calculated density ofabout 1.485 g cm⁻³; and b) increasing skin pigmentation in the subject.13. A method of treating cancer in a subject in need thereof comprising:administering to a subject a therapeutically effective amount of acompound of the formula:

1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-1-oneSRR G-1 or a pharmaceutically acceptable salt thereof, wherein thecompound comprises a crystalline Form A characterized by an XRPD patternhaving peaks expressed in degrees 2θ (±0.20) at about 5.75, about 20.54,about 20.71, about 21.25, and about 21.86, wherein the crystalline FormA has a melt onset of about 176° C., wherein the crystalline Form A isanhydrous, wherein the crystalline Form A exhibits low hygroscopicity,and wherein the crystalline Form A exhibits a calculated density ofabout 1.485 g cm⁻³; and b) alleviating one or more symptoms associatedwith the cancer in the subject.